U.S. patent application number 10/132677 was filed with the patent office on 2002-12-19 for 1-(n-phenylalkylaminoalkyl)piperazine derivatives substituted at position 2 of the phenyl ring.
This patent application is currently assigned to Recordati S.A., Chemical and Pharmaceutical Comoany. Invention is credited to Leonardi, Amedeo, Motta, Gianni, Riva, Carlo, Testa, Rodolfo.
Application Number | 20020193383 10/132677 |
Document ID | / |
Family ID | 27453039 |
Filed Date | 2002-12-19 |
United States Patent
Application |
20020193383 |
Kind Code |
A1 |
Leonardi, Amedeo ; et
al. |
December 19, 2002 |
1-(N-phenylalkylaminoalkyl)piperazine derivatives substituted at
position 2 of the phenyl ring
Abstract
The present invention is directed to novel
1-(N-phenylaminoalkyl)piperazin- e derivatives substituted at the
position 2 of the phenyl ring. Pharmaceutical compositions
comprising the compounds of the invention also are contemplated.
The compounds of the present invention also are contemplated for
use in treating neuromuscular dysfunction of the lower urinary
tract in a mammal.
Inventors: |
Leonardi, Amedeo; (Milano,
IT) ; Motta, Gianni; (Barlassina, IT) ; Riva,
Carlo; (Varese, IT) ; Testa, Rodolfo;
(Vignate, IT) |
Correspondence
Address: |
DARBY & DARBY P.C.
805 Third Avenue
New York
NY
10022
US
|
Assignee: |
Recordati S.A., Chemical and
Pharmaceutical Comoany
|
Family ID: |
27453039 |
Appl. No.: |
10/132677 |
Filed: |
April 22, 2002 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10132677 |
Apr 22, 2002 |
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09532505 |
Mar 21, 2000 |
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6399614 |
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09532505 |
Mar 21, 2000 |
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09127057 |
Jul 31, 1998 |
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6071920 |
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60070268 |
Dec 31, 1997 |
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Current U.S.
Class: |
514/253.13 ;
514/255.03; 544/360; 544/392 |
Current CPC
Class: |
C07D 241/24 20130101;
C07D 209/08 20130101; C07D 307/68 20130101; C07D 213/82 20130101;
C07D 213/81 20130101; C07D 295/24 20130101; C07D 333/38 20130101;
C07D 295/13 20130101 |
Class at
Publication: |
514/253.13 ;
514/255.03; 544/360; 544/392 |
International
Class: |
A61K 031/496; A61K
031/495; C07D 43/02; C07D 241/04 |
Foreign Application Data
Date |
Code |
Application Number |
Aug 1, 1997 |
IT |
MI97 001864 |
Claims
What is claimed:
1. A compound of the formula 8wherein R=is a hydrogen atom, an
alkylcarbonyl, a cycloalkylcarbonyl, a substituted
cycloalkylcarbonyl or a monocyclic heteroarylcarbonyl group;
R.sub.1 is chosen from the group consisting of hydrogen atom and
lower alkyl group; R.sub.2 is chosen from the group consisting of
halogen atom, alkoxy, phenoxy, nitro, cyano, acyl, amino,
acylamino, alkylsulphonylamino, alkoxycarbonyl,
N-acylaminocarbonyl, N-alkylaminocarbonyl,
N,N-dialkylaminocarbonyl, aminocarbonyl, trifluoromethyl and
polyfluroalkoxy groups; R.sub.3 is chosen from the group consisting
of methoxy and polyhaloalkoxy groups; R.sub.4 is chosen from the
group consisting of halogen aton, hydroxyl, lower alkoxy, lower
acyloxy, lower N-alkylaminocarbonyloxy and lower N,
N-dialkylaminocarbonyloxy groups; and n=1 or 2; or an enantiomer,
N-oxide, hydrate, or pharmaceutically acceptable salt thereof.
2. A compound according to claim 1 wherein n=1.
3. A compound according to claim 1 wherein R is
cycloalkylcarbonyl.
4. A compound according to claim 3 wherein R is
cyclohexylcarbonyl.
5. A compound according to claim 1 wherein R.sub.2 is selected from
the group consisting of alkoxy and polyfluroalkoxy groups.
6. A compound according to claim 5 wherein R.sub.2 is selected from
the group consisting of methoxy and trifluoromethoxy groups.
7. A compound according to claim 4 wherein R.sub.2 is selected from
the group consisting of methoxy and trifluoromethoxy groups.
8. A compound according to claim 1 wherein R.sub.3 is
polyhaloalkoxy and R.sub.4 is halogen.
9. A compound according to claim 8 wherein R is
cycloalkylcarbonyl.
10. A compound according to claim 9 wherein R is
cyclohexylcarbonyl.
11. A compound according to claim 8 wherein R.sub.2 is selected
from the group consisting of alkoxy and polyfluroalkoxy groups.
12. A compound according to claim 1 wherein R.sub.4 is chosen from
the group consisting of hydroxyl, lower alkoxy, lower acyloxy,
lower N-alkylaminocarbonyloxy and lower N,
N-dialkylaminocarbonyloxy groups.
13. A compound according to claim 12 wherein R.sub.4 is chosen from
the group consisting of lower acyloxy, lower
N-alkylaminocarbonyloxy and lower N, N-dialkylaminocarbonyloxy
groups.
14. A compound according to claim 13 wherein R is
cycloalkylcarbonyl.
15. A compound according to claim 14 wherein R is
cyclohexylcarbonyl.
16. A compound according to claim 15 wherein R.sub.2 is a
polyfluoroalkoxy group.
17. A compound according to claim 16 wherein R.sub.2 is a
trifluoromethoxy group.
18. A pharmaceutical composition comprising a compound of claim 1
and a pharmaceutically acceptable diluent, excipient or
carrier.
19. The pharmaceutical composition of claim 18 which comprises at
least one excipient selected from the group consisting of
lubricants, plasticizers, colorants, absorption enhancers, and
bactericides.
20. A compound selected from the group consisting of:
1-[N-cyclohexylcarbonyl-N-(2-methoxyphenyl)-2-aminoethyl]-4-[4-fluoro-2-(-
2,2,2-trifluoroethoxy)phenyl]piperazine;
1-[N-cyclohexylcarbonyl-N-(2-trif-
luoromethoxyphenyl)-2-aminoethyl]-4-[4-fluoro-2-(2,2,2-trifluoroethoxy)phe-
nyl]piperazine;
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-ami-
noethyl]-4-(2,4-dimethoxyphenyl)piperazine;
1-[N-cyclohexylcarbonyl-N-(2-t-
rifluoromethoxyphenyl)-2-aminoethyl]-4-(4-hydroxy-2-methoxyphenyl)piperazi-
ne;
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]-4-(-
4-acetoxy-2-methoxyphenyl)piperazine; 1
-[N-cyclohexylcarbonyl-N-(2-triflu-
oromethoxyphenyl)-2-aminoethyl]-4-(4-ethylaminocarbonyloxy-2-methoxyphenyl-
)piperazine; and
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-am-
inoethyl]-4-[4-(2-methylpropionyloxy)-2-methoxyphenyl]piperazine;
and enantiomers, N-oxides, hydrates, and pharmaceutically
acceptable salts thereof.
21. A pharmaceutical composition comprising a compound of claim 20
and a pharmaceutically acceptable diluent, excipient or
carrier.
22. The pharmaceutical composition of claim 21 which comprises at
least one excipient selected from the group consisting of
lubricants, plasticizers, colorants, absorption enhancers, and
bactericides.
23. A method for treating neuromuscular dysfunction of the lower
urinary tract in a mammal in need of such treatment, said method
comprising administering to said mammal an effective amount for
treating said dysfunction a compound of compound of the formula
9wherein R=is a hydrogen atom, an alkylcarbonyl, a
cycloalkylcarbonyl, a substituted cycloalkylcarbonyl or a
monocyclic heteroarylcarbonyl group; R.sub.1 is chosen from the
group consisting of hydrogen atom and lower alkyl group; R.sub.2 is
chosen from the group consisting of halogen atom, alkoxy, phenoxy,
nitro, cyano, acyl, amino, acylamino, alkylsulphonylamino,
alkoxycarbonyl, N-acylaminocarbonyl, N-alkylaminocarbonyl,
N,N-dialkylaminocarbonyl, aminocarbonyl, trifluoromethyl and
polyfluroalkoxy groups; R.sub.3 is chosen from the group consisting
of methoxy and polyhaloalkoxy; R.sub.4 is chosen from the group
consisting of halogen, hydroxyl, lower alkoxy, lower acyloxy, lower
N-alkylaminocarbonyloxy and lower N, N-alkylaminocarbonyloxy; and
n=1 or 2; or an enantiomer, N-oxide, hydrate or pharmaceutically
acceptable salt thereof.
24. The method of claim 23 wherein R is a cycloalkylcarbonyl group,
R.sub.2 is an alkoxy or trifluoroalkoxy group, R.sub.3 is a
polyhaloalkoxy group and R.sub.4 is a halogen atom.
25. The method of claim 23 wherein R is a cycloalkylcarbonyl group,
R.sub.2 is a trifluoroalkoxy group, R.sub.3 is a methoxy group and
R.sub.4 is selected from the group consisting of hydroxyl, lower
alkoxy, lower acyloxy, lower N-alkylaminocarbonyloxy and lower N,
N-dialkylaminocarbonyloxy groups.
26. The method of claim 23 wherein said administration is effective
for ameliorating at least one of urinary urgency, increased urinary
frequency, incontinence, urine leakage, enuresis, dysuria, urinary
hesitancy, and difficulty in bladder emptying in said mammal.
27. The method of claim 26 wherein said mammal is a human.
28. The method of claim 23, wherein said administering is achieved
using a route selected from the group consisting of oral, enteral,
intravenous, intramuscular, subcutaneous, transmucosal,
transdermal, and by-inhalation routes.
29. The method of claim 23, wherein said compound is administered
to said mammal in an amount of between about 0.01 and 25
mg/kg/day.
30. The method of claim 23, wherein said compound is administered
to said mammal in an amount of between about 0.2 and about 5
mg/kg/day.
31. The method of claim 23, wherein said compound is administered
to said mammal in an amount of between about 50 and 400 mg/day.
32. The method of claim 28, wherein said administering is achieved
using a route selected from the group consisting of oral and
transdermal routes.
33. The method of claim 32, wherein the amount of said compound is
between about 0.1 and 10 mg/kg/day.
34. A method for treating neuromuscular dysfunction of the lower
urinary tract in a mammal in need of such treatment, said method
comprising administering to said mammal an effective amount for
treating said dysfunction a compound chosen from the group
consisting
1-[N-cyclohexylcarbonyl-N-(2-methoxyphenyl)-2-aminoethyl]-4-[4-fluoro-2-(-
2,2,2-trifluoroethoxy)phenyl]piperazine;
1-[N-cyclohexylcarbonyl-N-(2-trif-
luoromethoxyphenyl)-2-aminoethyl]-4-[4-fluoro-2-(2,2,2-trifluoroethoxy)phe-
nyl]piperazine;
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-ami-
noethyl]-4-(2,4-dimethoxyphenyl)piperazine;
1-[N-cyclohexylcarbonyl-N-(2-t-
rifluoromethoxyphenyl)-2-aminoethyl]-4-(4-hydroxy-2-methoxyphenyl)piperazi-
ne;
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]-4-(-
4-acetoxy-2-methoxyphenyl)piperazine;
1-[N-cyclohexylcarbonyl-N-(2-trifluo-
romethoxyphenyl)-2-aminoethyl]-4-(4-ethylaminocarbonyloxy-2-methoxyphenyl)-
piperazine; and
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-ami-
noethyl]-4-[4-(2-methylpropionyloxy)-2-methoxyphenyl]piperazine;
and enantiomers, N-oxides, hydrates, and pharmaceutically
acceptable salts thereof.
35. The method of claim 34, wherein said administration is
effective for ameliorating at least one of urinary urgency,
increased urinary frequency, incontinence, urine leakage,
entiresis, dysuria, urinary hesitance, and difficulty in bladder
emptying in said mammal.
36. The method of claim 35 wherein said mammal is a human.
37. The method of claim 34, wherein said administering is achieved
using a route selected from the group consisting of oral, enteral,
intravenous, intramuscular, subcutaneous, transmucosal,
transdermal, and by-inhalation routes.
38. The method of claim 34, wherein said compound is administered
to said mammal in an amount of between about 0.01 and 25
mg/kg/day.
39. The method of claim 34, said compound is administered to said
mammal in an amount of between about 0.2 and about 5 mg/kg/day.
40. The method of claim 34, wherein said compound is administered
to said mammal in an amount of between about 50 and 400 mg/day.
41. The method of claim 34, wherein said administering is achieved
using a route selected from the group consisting of oral and
transdermal routes.
42. The method of claim 41, wherein the amount of said compound is
between about 0.1 and 10 mg/kg/day.
Description
RELATED APPLICATIONS
[0001] This application is a continuation-in-part of Ser. No.
09/532,505, filed Mar. 21, 2000, which is a continuation-in-part of
Ser. No. 09/127,057, filed Jul. 31, 1998, now U.S. Pat. No.
6,071,920, which claims priority under 35 U.S.C. .sctn.119 (e) of
U.S. Provisional Patent Application Serial No. 60/070,268, filed
Dec. 31, 1997 and priority under 35 U.S.C. .sctn.119 (b) of Italian
Patent Application No. M197 001864, filed Aug. 1, 1997. Each of the
aforementioned applications and patents is hereby incorporated by
reference herein in its entirety.
FIELD OF THE INVENTION
[0002] This invention relates to 1-(N-phenylaminoalkyl)piperazine
derivatives substituted at position 2 of the phenyl ring, to
pharmaceutical compositions containing them and to uses for such
derivatives and compositions .
BACKGROUND OF THE INVENTION
[0003] In mammals, micturition (urination) is a complex process
that requires the integrated actions of the bladder, its internal
and external sphincters, the musculature of the pelvic floor, and
neurological control over these muscles at three levels (in the
bladder wall or sphincter itself, in the autonomic centers of the
spinal cord, and in the central nervous system at the level of the
pontine micturition center (PMC) in the brainstem (pons) under the
control of cerebral cortex) (De Groat, Neurobiology of
Incontinence, (Ciba Foundation Symposium 151:27, 1990). Micturition
results from contraction of the detrusor muscle, which consists of
interlacing smooth muscle fibers under parasympathetic autonomic
control from the sacral spinal cord. A simple voiding reflex is
formed by sensory nerves for pain, temperature, and distension that
run from the bladder to the sacral cord. However, sensory tracts
from the bladder also reach the PMC, resulting in the generation of
nerve impulses that normally suppress the sacral spinal reflex arc
controlling bladder emptying. Thus, normal micturition is initiated
by voluntary suppression of cortical inhibition of the reflex arc
and by relaxation of the muscles of the pelvic floor and the
external sphincter. Finally, the detrusor muscle contracts and
voiding occurs.
[0004] Abnormalities of lower urinary tract function, e.g.,
dysuria, incontinence, and enuresis, are common in the general
population. Dysuria includes urinary frequency, nocturia, and
urgency, and may be caused by cystitis, prostatitis or benign
prostatic hypertrophy (BPH) (which affects about 70% of elderly
males), or by neurological disorders. Incontinence syndromes
include stress incontinence, urgency incontinence, and overflow
incontinence. Enuresis refers to the involuntary passage of urine
at night or during sleep.
[0005] Prior to the work of the present inventors, treatment of
neuromuscular dysfunction of the lower urinary tract has involved
administration of compounds that act directly on the bladder
muscles, such as flavoxate, a spasmolytic drug (Ruffinan, J. Int.
Med. Res. 16:317, 1988) also active on the PMC (Guarneri et al.,
Drugs of Today 30:91, 1994), or anticholinergic compounds such as
oxybutynin (Andersson, Drugs 35:477, 1988). The use of
.alpha..sub.1-adrenergic receptor antagonists for the treatment of
BPH is also common but is based on a different mechanism of action.
(Lepor, Urology, 42:483, 1993).
[0006] However, treatments that involve direct inhibition of the
pelvic musculature (including the detrusor muscle) may have
unwanted side effects such as incomplete voiding or accommodation
paralysis, tachycardia and dry mouth (Andersson, Drugs 35:477,
1988). Thus, it would be advantageous if compounds were available
that act via the peripheral or central nervous system to, for
example, affect the sacral spinal reflex arc and/or the PMC
inhibition pathways in a manner that restores normal functioning of
the micturition mechanism.
[0007]
1-(N-phenyl-N-cyclohexylcarbonyl-2-aminoethyl)-4-(2-methoxyphenyl)p-
iperazine (compound A) 1
[0008] is described in GB 2 263110 A and is reported to be a
5-HT.sub.1A receptor antagonist. It is also disclosed that it can
be used for the treatment of central nervous system disorders, for
example as an anxiolytic agent in the treatment of anxiety.
[0009] The compounds of the present invention, described below, are
structurally different from compound A because of the novel
substituents present on the aniline ring at the 2 position. Other
differences between the compounds of the present invention and
those disclosed in GB 2 263110 A are the substitutions on the
aromatic ring at position 4 of the piperazine ring. These
structural variations are neither disclosed nor suggested by GB 2
263110 A, particularly with regard to compounds that can be used to
improve urinary tract function. These structural variations result
in compounds that are more potent than compound A in
pharmacological tests predictive of activity on the lower urinary
tract, in particular for activity against urinary incontinence.
[0010] Other compounds which have been found by the present
inventors to be useful in the methods of the present invention,
e.g., treatment of disorders of the urinary tract, are disclosed in
U.S. Pat. No. 4,205,173; EP 711,757; DE patent 2,405,441; Chem.
Pharm. Bull. 33:1823-1835 (1985), and J. Med. Chem. 7:721-725
(1964), all of which are incorporated by reference.
SUMMARY OF THE INVENTION
[0011] In one aspect the invention is directed to compounds of
formula I: 2
[0012] wherein
[0013] R is hydrogen
[0014] R.sub.1 is chosen from the group consisting of hydrogen and
lower alkyl;
[0015] R.sub.2 is chosen from the group consisting of alkoxy,
phenoxy, nitro, cyano, acyl, amino, acylamino, alkylsulphonylamino,
alkoxycarbonyl, N-acylaminocarbonyl, N-alkylaminocarbonyl,
N,N-dialkylaminocarbonyl, aminocarbonyl, halo, trifluoromethyl or
polyfluroalkoxy group;
[0016] n=1 or 2;
[0017] B is chosen from the group consisting of optionally
substituted aryl, optionally substituted bicyclic aryl group,
optionally substituted 9-member bicyclic heteroaromatic containing
one heteroatom, and an optionally substituted benzyl group,
[0018] with the proviso that if B is aryl and is substituted by an
alkoxy group, the alkoxy group must be at the two position; and
[0019] if R.sub.1 is hydrogen and R.sub.2 is a nitro group and n=1,
then B cannot be a phenyl, 2 methoxyphenyl 4-chlorophenyl, 3
acetylphenyl, 3,4,5 trimethoxyphenyl , 2-chloro-4-methylphenyl or
2-pyridyl group; and
[0020] if R and R.sub.1 are hydrogen and B is optionally
substituted phenyl, then R.sub.2 cannot be acyl, acylamino,
alkoxycarbonyl, N-acylaminocarbonyl, N-alkylaminocarbonyl,
N,N-dialkylaminocarbonyl;
[0021] and enantiomers N-oxides hydrates and pharmaceutically
acceptable salts thereof.
[0022] Preferred is when n=1. Further preferred is when B is
optionally substituted phenyl. Further preferred is when B is
indolyl.
[0023] Also preferred is when R.sub.1 is hydrogen; R.sub.2 is
chosen from the group consisting of alkoxy, phenoxy, nitro, cyano,
amino, halo, trifluoromethyl or polyfluroalkoxy; n=1; and B is
substituted phenyl.
[0024] Also preferred is when R.sub.1 is hydrogen; R.sub.2 is
chosen from the group consisting of alkoxy, phenoxy, nitro, cyano,
amino, halo, trifluoromethyl or polyfluroalkoxy; n=1; and B is
indolyl.
[0025] Also preferred is when R.sub.1 is hydrogen; R.sub.2 is
chosen from the group consisting of alkoxy, nitro, halo,
trifluoromethyl or polyfluroalkoxy; n=1; and B is substituted
phenyl.
[0026] Also preferred is when R.sub.1 is hydrogen; R.sub.2 is
chosen from the group consisting of alkoxy, nitro, halo,
trifluoromethyl or polyfluroalkoxy; n=1; and B is substituted
indolyl.
[0027] In yet another aspect, the invention is directed to
compounds of formula I B 3
[0028] wherein
[0029] R is a hydrogen atom, an alkylcarbonyl, a
cycloalkylcarbonyl, a substituted cycloalkylcarbonyl or a
monocyclic heteroarylcarbonyl group,
[0030] n=1 or2;
[0031] R.sub.1 is a hydrogen atom or a lower alkyl group,
[0032] R.sub.2 is an alkoxy, phenoxy, nitro, cyano, acyl, amino,
acylamino, alkylsulphonylamino, alkoxycarbonyl, aminocarbonyl,
N-alkylaminocarbonyl, N,N-dialkylaminocarbonyl,
N-acylaminocarbonyl, halo, trifluoromethyl or polyfluoroalkoxy
group; and
[0033] B is a bicyclic heteroaromatic with the proviso that B is
not a 9-member bicyclic heteroaromatic containing one
heteroatom.
[0034] Preferred is when R is chosen from the group consisting of
hydrogen and cycloalkylcarbonyl; R.sub.1 is hydrogen; R.sub.2 is
chosen from the group consisting of alkoxy, phenoxy, nitro, cyano,
acyl, amino, acylamino, alkylsulphonylamino, alkoxycarbonyl,
aminocarbonyl, N-alkylaminocarbonyl, N,N-dialkylaminocarbonyl,
N-acylaminocarbonyl, halo, trifluromethyl or polyfluroalkoxy group;
n=1; and B is a 10-member bicyclic heteroaromatic containing one
heteroatom.
[0035] Further preferred is when n=1.
[0036] Further preferred is when B is a 10-member bicyclic
heteroaromatic containing one heteroatom.
[0037] Further preferred is when B is an optionally substituted
quinolyl.
[0038] The invention also includes the enantiomers, diastereomers,
N-oxides, crystalline forms, hydrates and pharmaceutically
acceptable salts of compounds of the formula 1 and 1B and formula
1C, as well as metabolites of these compounds having the same type
of activity (hereafter sometimes referred to as "active
metabolites").
[0039] In a preferred aspect are provided compounds of formula 1C
4
[0040] wherein
[0041] R=is a hydrogen atom, an alkylcarbonyl, a
cycloalkylcarbonyl, a substituted cycloalkylcarbonyl or a
monocyclic heteroarylcarbonyl group;
[0042] R.sub.1 is chosen from the group consisting of hydrogen and
lower alkyl;
[0043] R.sub.2 is chosen from the group consisting of alkoxy,
phenoxy, nitro, cyano, acyl, amino, acylamino, alkylsulphonylamino,
alkoxycarbonyl, N-acylaminocarbonyl, N-alkylaminocarbonyl,
N,N-dialkylaminocarbonyl, aminocarbonyl, halo, trifluoromethyl or
polyfluroalkoxy group;
[0044] R.sub.3 is chosen from the group consisting of methoxy and
polyhaloalkoxy;
[0045] R.sub.4 is chosen from the group consisting of halogen,
hydroxyl, lower alkoxy, lower acyloxy, lower
N-alkylaminocarbonyloxy and lower N, N-dialkylaminocarbonyloxy;
and
[0046] n=1 or2;
[0047] and enantiomers, N-oxides, hydrates, and pharmaceutically
acceptable salts thereof.
[0048] Preferred compounds of formula 1C are where, independently,
R is a cycloalkylcarbonyl group, R.sub.1 is a hydrogen atom or
lower alkyl group, R.sub.2 is an alkoxy or trifluoromethyl group,
R.sub.3 is a methoxy or polyhaloalkoxy group, R.sub.4 is chosen
from the group consisting of halogen, hydroxyl, lower alkoxy, lower
acyloxy, lower N-alkylaminocarbonyloxy and lower N,
N-alkylarninocarbonyloxy, and n=1 or 2. With further respect to
compounds of formula 1C, a preferred cycloalkylcarbonyl group for R
is cyclohexylcarbonyl group, a preferred alkoxy group for R.sub.2
is methoxy, a preferred polyhaloalkoxy group for R.sub.3 is
2,2,2-trifluoroethoxy, a preferred lower alkoxy group for R.sub.4
is methoxy, preferred lower acyloxy groups for R.sub.4 are acetoxy
and 2-methylpropionyloxy, a preferred lower N-alkylaminocarbonyloxy
group for R.sub.4 is N-ethylaminocarbonyloxy and the preferred
value for n is 1. Each of the foregoing preferences are independent
of each other.
[0049] Further preferred are compounds of Formula 1C wherein,
simultaneously, R is cycloalkylcarbonyl, R.sub.2 is alkoxy or
polyfluoroalkoxy, R.sub.3 is polyfluoroalkoxy, R.sub.4 is halogen
and n=1 or 2. Also preferred are compounds of Formula 1C wherein,
simultaneously, R is cycloalkylcarbonyl, R.sub.2 is
polyfluoroalkoxy, R.sub.3 is methoxy, R.sub.4 is selected from the
group consisting of hydroxyl, lower alkoxy, lower acyloxy, lower
N-alkylaminocarbonyloxy and lower N, N-alkylaminocarbonyloxy
groups, and n=1 or 2.
[0050] In another aspect, the invention is directed to compounds of
formula 1C chosen from the group consisting of:
[0051]
1-[N-cyclohexylcarbonyl-N-(2-methoxyphenyl)-2-aminoethyl]-4-[4-fluo-
ro-2-(2,2,2-trifluoroethoxy)phenyl]piperazine;
[0052]
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]--
4-[4-fluoro-2-(2,2,2-trifluoroethoxy)phenyl]piperazine;
[0053]
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]--
4-(2,4-dimethoxyphenyl)piperazine;
[0054] 1
-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]-
-4-(4-hydroxy-2-methoxyphenyl)piperazine;
[0055]
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]--
4-(4-acetoxy-2-methoxyphenyl)piperazine;
[0056]
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]--
4-(4-ethylaminocarbonyloxy-2-methoxyphenyl)piperazine; and
[0057] 1-[N-cyclohexylcarbonyl-N-(2-
trifluoromethoxyphenyl)-2-aminoethyl]-
-4-[4-(2-methylpropionyloxy)-2-methoxyphenyl]piperazine;
[0058] and enantiomers, N-oxides, hydrates, and pharmaceutically
acceptable salts thereof.
[0059] As used herein with reference to variable R, alkylcarbonyl
radicals include C.sub.1-C.sub.6 alkylcarbonyl, cycloalkylcarbonyl
includes cyclohexylcarbonyl, substituted cycloalkylcarbonyl
includes cyclohexylcarbony substituted with alkyl or aryl groups an
d monocyclic heteroaryl radicals include monocyclic aromatic
radicals of 5 to 7 ring atoms containing one or more hetero atoms
(e.g., oxygen, nitrogen, and sulfur). Monocyclic heteroarylcarbonyl
has the same definition as monocyclic heteroaryl, but also
comprises a carbonyl group linked to a carbon atom of the ring.
[0060] As used herein with reference to variable B, a mono or
bicyclic aryl radical means an aromatic radical having 6 to 12
carbon atoms (e.g., phenyl or naphthyl) which is substituted by one
or more substitutents. Preferred substitutents for aryl radicals
are lower alkyl, hydroxy, lower acyloxy (e.g., acetoxy), lower
alkylaminocarbonyloxy (e.g., N, ethylaminocarbonyloxy and
2-methylpropionyloxy), lower alkoxy (e.g., methoxy, ethoxy,
propoxy, and butoxy), lower haloalkoxy (e.g.,
2,2,2-trifluoroethoxy) halogen, amino, acylamino,
alkylsulfonylamino, and (lower) alkylamino substituents.
[0061] As used with respect to variable B, monocyclic heteroaryl
radical has the same meaning as for R, above, and bicyclic
heteroaryl radical means a bicyclic aromatic radical containing one
or more heteroatoms (e.g., nitrogen, oxygen, sulfur) and 9 to 12
ring atoms. Benzyl radicals, with respect to variable B, include
phenylmethyl radicals which may be optionally substituted by one or
more substituents. Preferred substituents for the benzyl radicals
are alkyl, alkoxy, halogen, nitro, cyano, amido, amino, alkylamino,
acylamino, alkylsulphonylamino or acyl substituents. Preferred
substituents at B are optionally substituted monocyclic aryl and
bicyclic heteroaryl. Most preferred substituents at B are
alkoxyphenyl and mononitrogen-containing bicyclic heteroaryl.
[0062] Preferred substituents at R.sub.1 are hydrogen and
methyl.
[0063] Preferred substituents at R.sub.2 are nitro, cyano, acyl,
alkoxy, trifluoralkoxy and aminocarbonyl. More preferred at R.sub.2
are nitro, alkoxy and trifluoralkoxy. A preferred value for n is
1.
[0064] Preferred substituents for B are optionally substituted
phenyl and indolyl.
[0065] The invention further provides pharmaceutical compositions
comprising a compound of formula I or a compound of formula 1B, or
a compound of formula 1C, or an enantiomer, diastereomer, N-oxide,
crystalline form, hydrate or pharmaceutically acceptable salt of
the compound, in admixture with a pharmaceutically acceptable
diluent or carrier.
[0066] In another aspect, the present invention is directed to
methods for reducing the frequency of bladder contractions due to
bladder distension by administering one or more selected compounds
of Formula I, or a compound of formula 1B, or a compound of formula
1C, to a mammal (including a human) in need of such treatment, in
an amount or amounts effective for the particular use.
[0067] In a further aspect, the present invention is directed to
methods for treating disorders of the urinary tract in a subject in
need of such treatment, comprising administering an effective
amount of a compound of Formula 1, or a compound of formula 1B, or
a compound of formula 1C, to ameliorate at least one of urinary
urgency, increased urinary frequency, incontinence, urine leakage,
enuresis, dysuria, urinary hesitancy, and difficulty in emptying
bladder.
[0068] In yet another aspect, the invention is directed to methods
for blocking 5-HT.sub.1A serotonergic receptors, and, by virtue of
this inhibitory activity, to methods for the treatment of CNS
disorders due to serotonergic dysfunction such as anxiety,
depression, hypertension, sleep/wake cycle disorders, feeding
behavior, sexual function and cognition disorders in mammals,
particularly in humans, by delivering to the environment of the
5-HT.sub.1A serotonergic receptors, e.g., to the extracellular
medium (or by administering to a mammal possessing such receptors)
an effective amount of a compound of formula 1, formula 1B or
formula 1C (hereinafter "compounds of the invention").
DETAILED DESCRIPTION OF THE INVENTION
[0069] All patents, patent applications, and literature references
cited in the specification are hereby incorporated by reference in
their entirety. In the case of inconsistencies, the present
disclosure, including definitions, will prevail.
[0070] The activity of the compounds of the invention as inhibitors
of frequency of micturition renders them useful for the treatment
of neuromuscular dysfunctions of the lower urinary tract in
mammals, including without limitation dysuria, incontinence and
enuresis. Surprisingly, the introduction of selected substituents
at position 2 of the aniline ring in compounds of Formula I,
Formula IB and Formula 1C confers upon these compounds a distinctly
higher potency with regard to compound A and also to the isomers
bearing the same substituent at position 3 or 4. This information
was obtained by testing compound A and the corresponding 2, 3 and 4
nitroaniline derivatives (Example 2 and compounds B and C) in a rat
model. The rhythmic contraction of rat bladders was induced by
filling the bladders with a physiologic solution. The effect of
test compounds of the invention on the frequency and amplitude of
the contractions was evaluated. Of particular interest is the time
of disappearance of induced contractions. (ED.sub.10 min in Table
1, below).
[0071] Data in Table 1 (in particular, ED.sub.50 (frequency)) show
that the 2-substituted compounds of the invention are clearly more
potent inhibitors of the frequency of urinary bladder
contractions.
[0072] The effect of the drugs currently available for
administration to humans for treatment of neuromuscular function of
the lower urinary tract (flavoxate and oxybutynin) on the
above-described rat model is also shown, for comparative purposes,
in Table 1.
[0073] The compounds of the invention are more potent and acted for
a longer period of time (as measured by duration of bladder
quiescence with no contractions) than did compounds A, flavoxate,
and oxybutynin. Moreover, in contrast to oxybutynin, the compounds
of the invention did not affect the amplitude of the contractions
(no effect on ED.sub.50 (amplitude) in Table 1), suggesting no
impairment of bladder contractility that could result in residual
urine being left in the bladder after micturition.
[0074] In addition, the beneficial effect on the lower urinary
tract of the compounds of the invention has been shown in a
cystometry model in conscious rats, where the compounds of the
invention are also superior to compound A and the reference drugs.
In fact, compounds of the invention increased the bladder capacity
at doses lower than compound A and flavoxate (oxybutynin does not
affect bladder capacity) (Table 2). Furthermore, in contrast to the
effects of oxybutynin, no impairment of bladder contractility
(decrease in MP) was observed.
[0075] Finally, the compounds of the invention have a high and
selective affinity for the 5-HT.sub.1A receptor, an affinity and
selectivity displayed to a much lesser extent by compound A, (Table
3). The compounds of the invention have also been shown to
antagonize both pre- and post-synaptic 5-HT.sub.1A receptors much
more potently than compound A (Table 4), strongly suggesting a role
for the 5-HT.sub.1A receptor in the action of the compounds of the
invention.
[0076] Subjects who can benefit from administration of the
compounds and compositions of the invention include humans who are
affected by neuromuscular dysfunction of the lower urinary tract,
described by E. J. McGuire in "Campbell's UROLOGY" 5.sup.th Ed.
616-638, 1986, W. B. Saunders Company, and also include patients
affected by any physiological dysfunction related to impairment of
5-HT.sub.1A receptor function. Such dysfunctions include, without
limitation, central nervous system disorders such as depression,
anxiety, eating disorders, sexual dysfunction, addiction, and
related problems.
[0077] The present invention encompasses pharmaceutical
formulations comprising the compounds disclosed above, as well as
methods employing these formulations for treating neuromuscular
dysfunction of the lower urinary tract such as dysuria,
incontinence, enuresis, and the like. Dysuria includes urinary
frequency, nocturia, urgency, and difficulty in emptying the
bladder, i.e., a suboptimal volume of urine is expelled during
micturition.
[0078] Incontinence syndromes include stress incontinence, urgency
incontinence, and overflow incontinence. Enuresis refers to the
involuntary passage of urine at night or during sleep.
[0079] Without wishing to be bound by theory, it is believed that
administration of the 5-HT.sub.1A receptor antagonists of the
invention prevents unwanted activity of the sacral reflex arc
and/or cortical mechanisms that control micturition. Thus it is
contemplated that a wide range of neuromuscular dysfunctions of the
lower urinary tract can be treated using the compounds of the
present invention.
[0080] An "effective amount" of the compound for treating a urinary
disorder is an amount that results in measurable amelioration of at
least one symptom or parameter of the disorders described
above.
[0081] An effective amount for treating the disorder can easily be
determined by empirical methods known to those of ordinary skill in
the art, such as by establishing a matrix of dosages and
frequencies of administration and comparing a group of experimental
units or subjects to each point in the matrix. The exact amount to
be administered to a patient will vary depending on the state and
severity of the disorder and the physical condition of the patient.
A measurable amelioration of any symptom or parameter can be
determined by a physician skilled in the art or reported by the
patient to the physician. It will be understood that any clinically
or statistically significant attenuation or amelioration of any
symptom or parameter of urinary tract disorders is within the scope
of the invention. Clinically significant attenuation or
amelioration means perceptible to the patient and/or to the
physician.
[0082] For example, a single patient may suffer from several
symptoms of dysuria simultaneously, such as, for example, urgency
and excessive frequency of urination, either or both of which may
be reduced using the methods of the present invention. In the case
of incontinence, any reduction in the frequency or volume of
unwanted passage of urine is considered a beneficial effect of the
present methods of treatment.
[0083] The compounds of the present invention may be formulated
into liquid dosage forms with a physiologically acceptable carrier,
such as, for example, phosphate buffered saline or deionized water.
The pharmaceutical formulation may also contain excipients,
including preservatives and stabilizers, that are well-known in the
art. The compounds can be formulated into solid oral or non-oral
dosage units such as, for example, tablets, capsules, powders, and
suppositories, and may additionally include excipients, including
without limitation lubricant(s), plasticizer(s), colorant(s),
absorption enhancer(s), bactericide(s), and the like.
[0084] Modes of administration include oral and enteral,
intravenous, intramuscular, subcutaneous, transdermal, transmucosal
(including rectal and buccal), and by-inhalation routes.
Preferably, an oral or transdermal route is used (i.e., via solid
or liquid oral formulations, or skin patches, respectively).
[0085] The amount of the agent to be administered can range from
between about 0.01 and about 25 mg/kg/day, preferably from between
about 0.1 and about 10 mg/kg/day and most preferably from between
about 0.2 and about 5 mg/kg/day. It will be understood that the
single pharmaceutical formulations of the present invention need
not contain the entire amount of the agent that is effective in
treating the disorder, as such effective amounts can be reached by
administration of a plurality of doses of such pharmaceutical
formulations.
[0086] In a preferred embodiment of the present invention,
compounds are formulated in capsules or tablets, each preferably
containing 50-200 mg of the compounds of the invention, and are
most preferably administered to a patient at a total daily dose of
50-400 mg, preferably 150-250 mg, and most preferably about 200 mg
for relief of urinary incontinence and dysfunctions amenable to
treatment with 5-HT.sub.1A receptor ligands.
[0087] The methods, tables and examples provided below are intended
to more fully describe preferred embodiments of the invention and
to demonstrate its advantages and applicability, without in any way
limiting the scope of the invention.
SYNTHESIS OF THE COMPOUNDS OF THE INVENTION
[0088] The compounds of the invention may be prepared by the
methods illustrated in the following reaction schemes, or by
modifications thereof, using readily available starting materials,
reagents and conventional synthesis procedures well known to those
of ordinary skill in the art.
[0089] Unless otherwise specified, the substituents of the
compounds and intermediates present in the reaction schemes are
defined in the same manner as they are defined above in formula I,
formula IB and formula 1C. One method to synthesize compounds of
formula I and formula IB (R=H) and formula IB and Formula 1C (R=H)
is depicted in Scheme 1.
[0090] Ortho-substituted anilines of formula II (Y=NH.sub.2) are
alkylated with 1,.omega.-disubstituted alkanes (Z) to give product
III. The reaction is carried out in an inert organic solvent,
preferentially a polar aprotic solvent such as
N,N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), dioxane,
tetrahydrofuran (THF), acetone, acetonitrile or chlorinated
solvents such as dichloromethane, chloroform, 1,2-dichloroethane or
a protic solvent such as n-butanol (n-BuOH). The reactions are
generally performed at a temperature between 0.degree. C. and
+120.degree. C., in the presence of a proton acceptor such as
triethylamine (Et.sub.3N), diisopropylethylamine, or the like, and
optionally in the presence of potassium iodide.
[0091] In compounds of formula Z, X and X.sub.1 can be Cl, Br, I,
aryl, or alkylsulfonyloxy 5
[0092] Intermediates of formula III are used in the alkylation of
suitable piperazine derivatives IV to give the compounds of formula
I and IB (where R=H).
[0093] These alkylations may be carried out in a chlorinated
solvent such as dichlorometane, chloroform or 1,2-dichloroethane,
or in a polar aprotic solvent such as DMF, THF, acetone,
acetonitrile, or in a polar protic solvent such as n-BuOH, etc., or
in an a polar solvent such as toluene, benzene, n-heptane, etc., at
a temperature between 0.degree. C. and 120.degree. C., optionally
in the presence of a proton acceptor, such as Et.sub.3N,
4-dimethylaminopyridine, potassium carbonate, cesium carbonate, and
the like, and optionally in the presence of potassium iodide.
[0094] Piperazines of formula IV which are not commercially
available may be prepared by reaction of the suitable B-NH.sub.2
derivatives (which generally may be easily obtained by reduction of
the corresponding B-NO.sub.2 derivatives) with
bis-(2-chloroethyl)amine or bis-(2-hydroxyethyl)amine in presence
of excess hydrogen chloride. These reactions can be performed in
aprotic solvents such as dimethylformamide, diglyme or toluene at a
temperature between +40.degree. C. and the reflux temperature of
the solvent, generally in the presence of a base such as potassium
carbonate, cesium carbonate, or the like, and optionally in the
presence of potassium iodide.
[0095] Compounds of formula V can be conveniently prepared starting
from compounds V in which X is a COO-lower alkyl group and n is
n-1. Conventional reduction procedures (e.g., use of lithium
aluminum hydride or other metal complex hydrides) afford the
corresponding compounds V in which X is CH.sub.2OH and n is n-1,
which can be in turn conventionally converted into compounds of
formula V in which X is a leaving group as described above. The
starting esters can be prepared by well known Michael reactions or
by the nucleophilic displacement reaction of a monosubstituted
piperazine on the appropriate 2,3-unsaturated ester or
2-haloester.
[0096] Alternative procedures to obtain compounds of formula V
consists in alkylating the appropriate monosubstituted piperazine
derivatives with a compound with the formula
X--CH(R.sub.1)(CH.sub.2).sub.n-1CH.sub.2--OPrG or
X--(CH.sub.2).sub.nCH(R.sub.1)--X where X is a leaving group and n
has the same meaning as above, and PrG is a protecting group (e.g.
O-tetrahydropyranyl), which can be removed after alkylation of the
piperazine.
[0097] Another approach to synthesize intermediate compounds of
formula III utilizes starting materials with structure II
(Y.dbd.halogen). These starting materials are reacted with
compounds of formula Z in which X and X.sub.1 are, respectively,
NH.sub.2 and OH. These alkylation reactions are carried out in an
aprotic solvent such as DMF, toluene, or in a polar protic solvent
such as n-BuOH, etc., at a temperature between +40.degree. C. and
+140.degree. C., in general using one equivalent or excess of a
reagent of formula Z (X.dbd.NH.sub.2) as a proton acceptor, as
described by G. Doleschall et al., Tetrahedron, 32, 57-64 (1976).
The resulting aminoalcohols of formula III (X.sub.1.dbd.OH) are
reacted with a chlorinating agent such as POCl.sub.3, SOCl.sub.2 or
PCl.sub.5 to give the intermediates, also of formula III
(X.sub.1.dbd.Cl), or with an alkyl or arylsulfonyl chloride to give
the corresponding sulfonyl esters. These reactions are carried out
in an aprotic solvent such as chloroform, DMF, pyridine, and the
like at a temperature between +50.degree. C. and the reflux
temperature of the solvent.
[0098] Compounds of formula I and IB (R.dbd.H) may also be obtained
by alkylation of compounds of formula II (Y.dbd.NH.sub.2) with
intermediates of formula V, in which B, R.sub.1 and n have the same
meanings as above and X is a halogen atom such as chlorine or
bromine, or a leaving group such as methanesulfonyloxy or
p-toluenesulfonyloxy groups.
[0099] These reactions may be carried out without solvent or in an
aprotic solvent such as dichloromethane, chloroform, DMF, THF,
acetone, acetonitrile or in a protic solvent such as n-butanol,
etc. at a temperature between 0.degree. C. and +160.degree. C.,
optionally in the presence of a proton acceptor, such as Et.sub.3N,
potassium carbonate, cesium carbonate, 4-dimethylaminopyridine and
the like, and optionally in the presence of potassium iodide.
[0100] Compounds of formula I and IB where R.sub.2 is CN can be
also obtained from the compounds of formula I and IB in which
R.sub.2 is CONH.sub.2 by dehydration reactions. P.sub.2O.sub.5,
PCl.sub.5, Ph.sub.3P, and the like may be used as dehydrating
agents (J. March, Advanced Organic Chemistry, IV Ed., page 1041,
Wiley Interscience, 1992). Dehydration reactions may be carried out
in a chlorinated solvent such as dichloromethane, chloroform,
carbon tetrachloride or in an aprotic solvent such as DMF, toluene,
etc. at a temperature between +40.degree. C. and the reflux
temperature of the solvent, optionally in the presence of a base
such as Et.sub.3N.
[0101] Alternatively,compounds of formula I and IB (R.dbd.H) may be
obtained by arylation of intermediates of formula V
(X.dbd.NH.sub.2) with a starting material of formula II (Y.dbd.Cl,
Br, F, I or trifluoromethanesulphonyloxy). These reactions may be
carried out using the same solvents and conditions as described
above or by employing palladium complex catalysis (Synlett,p.329
(1996)).
[0102] Compounds of formula I and IB in which R.sub.2 is COalk can
be synthesized from compounds I in which R.sub.2 is H by an
acylation reaction that can be carried out using boron trichloride
as a Lewis acid and acetonitrile as a reagent in an aprotic solvent
such as chloroform, 1,2-dichloroethane, toluene, etc. at
temperatures between 0.degree. C. and 100.degree. C., followed by
acidic hydrolysis by treatment with HCl at 100.degree. C., (T.
Sugasawa et al., Chem. Pharm. Bull., 33, 1826-1835 (1985)).
[0103] Compounds of formula I and IB (R.dbd.H) are acylated to give
compound IB (R other than H) by reaction with an appropriate acyl
halide R'Hal in which R' represents an alkylcarbonyl,
cycloalkylcarbonyl or monocyclic heteroarylcarbonyl group and Hal
represents a halogen atom. The reaction can be performed in aprotic
solvents such as dichloromethane, chloroform, 1,2-dichloroethane,
DMF, acetone, acetonitrile, toluene, etc., at temperatures between
0.degree. C. and 100.degree. C., optionally in the presence of an
organic base as a proton acceptor such as Et.sub.3N,
diisopropylethylamine (DIPEA), 4-dimethylaminopyridine, and the
like.
[0104] Alternatively,compounds with formula IB (i.e, where
R.sub.2.dbd.Br, I, OSO.sub.2F or OSO.sub.2CF.sub.3) in which R is
as defined above, but is not hydrogen, may be used to synthesize
compounds of formula I in which R.sub.2 is CN, CONH.sub.2,
COCH.sub.3 or COOCH.sub.3 by reaction of reagents such as
trimethylsilyl isocyanate and t-butyl lithium (J. Org. Chem. 55,
3114 (1990)), lithium cyanide and tetrakis(triphenylphosphine)p-
alladium(0) (EP711757), carbon monoxide-methanol and palladium
diacetate in the presence of 1,3-diphenylphosphinopropane (J. Org.
Chem. 59, 6683 (1994)). Such reactions may be carried out in polar
or a polar solvent such as THF, toluene, benzene, DMSO, and the
like.
[0105] Another method to synthesize compounds of formula I and IB
in which R.sub.1 is H is depicted in Scheme 2, below. 6
[0106] Ortho-substituted halobenzenes of formula II (Y.dbd.halo)
are used to arylate protected aminoalkylaldehydes of formula VII
(X.dbd.NH.sub.2) to give the corresponding protected
arylaminoalkylaldehydes VIII. The reaction may be carried out in an
aprotic solvent such as pyridine, DMF, toluene, or the like at a
temperature between +40.degree. C. and 120.degree. C., optionally
in the presence of a base such as Et.sub.3N or employing palladium
complex catalysts as above.
[0107] Another route for the preparation of intermediates of
formula VIII consists in alkylating compounds of formula II
(Y.dbd.NH.sub.2) with protected reactive compounds of formula VII
(X.dbd.halo) by conventional procedures known to those skilled in
the art. Compounds with formula VIII are stable and are deprotected
by standard methods just before their use in the following
steps.
[0108] Aldehydes of formula VIII', obtained from deprotection of
compounds with formula VIII, may be reacted without isolation with
N-substituted piperazines IV under reductive conditions to give
compounds of formula I and IB (R.dbd.R.sub.1.dbd.H). These
reactions may be carried out in polar solvents such as methanol,
ethanol or in chlorinated solvents, such as dichloromethane,
chloroform, and the like, using alkali borohydrides such as
NaBH.sub.4 and NaBH.sub.3CN, NaBH(OAc).sub.3 or using borane
complexes such as BH.sub.3-Py, optionally in the presence of acidic
promoter, such as acetic acid, at temperatures between +10.degree.
C. and 100.degree. C.
[0109] Alternatively, intermediates of formula VIII may be acylated
with R'Hal to give compounds of formula IX using the same
conditions as described above.
[0110] Intermediates of formula IX are deprotected by well-known
methods just before their use in the final step to give the
corresponding aldehydes (IX'), which may be reacted with
appropriate N-substituted piperazines of formula IV using alkali
borohydrides such as NaBH.sub.4, NaBH.sub.3CN or NaBH(OAc).sub.3,
optionally in the presence of catalytic amounts of acetic acid, or
of a titanium catalyst such as titanium tetraisopropoxide, yielding
compounds of formula I. These reactions may be carried out in
chlorinated solvents such as dichloromethane or chloroform, or in
polar aprotic solvents such as methanol or ethanol at temperatures
between +10.degree. C. and +100.degree. C.
[0111] An alternative procedure to afford compounds of formula I,
IB and 1C (Scheme 3), where R.sub.2 is an electron withdrawing
group (i.e. NO.sub.2, CN, I) consists of acylating by conventional
procedure with R'-Hal (R'.dbd.alkylcarbonyl) the proper aniline
(II, Y.dbd.NH.sub.2) to obtain compounds X, which in turn can be
alkylated with compounds V where X is a leaving group. The
alkylation of compound X can be carried out by preforming the
aza-anion of X through the use of bases (e.g. potassium
tert-butoxide, NaNH.sub.2, Na, NaH, buthyl lithium or other lithium
bases, NaOH/KOH by phase transfer catalysis) in a solvent such as
toluene, dimethylsulfoxide, DMF, acetonitrile, acetone, diethyl
ether, dioxane, tetrahydrofurane at a temperature included in the
range between -25.degree. C. and the temperature of reflux of the
solvent. The following alkylation to afford compounds I and IB can
be performed by adding to the reaction mixture the compounds V in
the same reaction condition as above.
[0112] The same alkylation reaction of compounds X can be carried
out using compound VII as an alternative methods to prepare
compounds IX.
[0113] The compounds of formula IB and (R.dbd.alkylcarbonyl,
R.sub.2.dbd.alkylCO) can be prepared from the proper compounds X
(R.sub.2.dbd.alkylCO) by protecting the keto group (e.g. as
1,3-dioxolanyl derivatives) by standard procedures, then by
alkylating the nitrogen of the amido group as described above.
Subsequent deprotection affords the desired compounds IB. 7
[0114] In addition, compounds of formula I where R is
alkylcarbonyl, cycloalkylcarbonyl or monocyclic heteroarylcarbonyl
group could also be synthesized using this methodology.
[0115] Compounds of formula I where B is disubstituted phenyl and
one of the substitutions is a lower acyloxy or a lower
N-alkylaminocarbonyloxy or lower N, N-dialkylaminocarbonyl-oxy
group (i.e., wherein the alkyl groups attached to the nitrogen atom
of the respective N-alkylaminocarbonyloxy or N,
N-dialkylaminocarbonyloxy groups are lower alkyl groups) can be
prepared starting from the corresponding compound of formula I in
which one of the two substituents is a hydroxy group. Reactions can
be carried out by standard procedures of esterification or addition
of a lower alky isocyanate, well known to those skilled in the
art.
Compound AA
[0116] 1-(N-phenyl-2-aminoethyl)-4-(2-methoxyphenyl)piperazine
Compounds B and D
[0117]
1-[N-(3-nitrophenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(2-methox-
yphenyl)piperazine (Compound B); and
[0118]
1-[N-(3-nitrophenyl)-2-aminoethyl]-4-(2-methoxyphenyl)piperazine
(Compound D)
Compounds C and E
[0119]
1-[N-(4-nitrophenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(2-methox-
yphenyl)piperazine (Compound C); and
[0120]
1-[N-(4-nitrophenyl)-2-aminoethyl]-4-(2-methoxyphenyl)piperazine
(Compound E)
[0121] Methods for synthesizing Examples 1-89 are described in U.S.
patent application Ser. No. 09/532,505, filed Mar. 21, 2000, now
U.S. Pat. No. ______, which is hereby incorporated herein in its
entirety.
EXAMPLE 1
[0122]
1-[N-(2-nitrophenyl)-2-aminoethyl]-4-(2-methoxyphenyl)piperazine
EXAMPLE 2
[0123] 1-[N -(2-
nitrophenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(2-meth-
oxyphenyl) piperazine
EXAMPLE 3
[0124]
1N-(2-trifluoromethoxyphenyl)-2-aminoethyl]-4-(2-methoxyphenyl)
piperazine
EXAMPLE 4
[0125] 1-[N
-(2-trifluoromethoxyphenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-
-4-(2-methoxyphenyl)piperazine
EXAMPLE 5
[0126] 1-[N-(2-phenoxyphenyl)-2-aminoethyl]-4-(2-methoxyphenyl)
piperazine
EXAMPLE 6
[0127] 1-[N
-(2-phenoxyphenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(2-met-
hoxy phenyl)piperazine
EXAMPLE 7
[0128] 1-[N-(2-iodophenyl)-2-aminoethyl]-4-(2-methoxyphenyl)
piperazine
EXAMPLE 8
[0129]
1-[N-(2-iodophenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(2-methoxy-
phenyl) piperazine
EXAMPLE 9
[0130]
1-[N-(2-aminocarbonylphenyl)-2-amininoethyl]-4-(2-methoxyphenyl)
piperazine
EXAMPLE 10
[0131] 1-[N-(2-cyanophenyl)-2-aminoethyl]-4-(2-methoxyphenyl)
piperazine
EXAMPLE 11
[0132] 1-[N-(2-acetylphenyl)-2-aminoethyl]-4-(2-methoxyphenyl)
piperazine
EXAMPLE 12
[0133]
1-[N-(2-nitrophenyl)-2-aminoethyl]-4-(4-indolyl)piperazine
EXAMPLE 13
[0134]
1-[N-(2-nitrophenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(4-indoly-
l) piperazine
EXAMPLE 14
[0135] 1-[N-(2-nitrophenyl)-2-aminoethyl)-4-(2,5-dichlorobenzyl)
piperazine
EXAMPLE 15
[0136]
1-[N-(2-nitrophenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(2,5-dich-
lorobenzyl) piperazine
EXAMPLE 16
[0137] 1-
[N-(2-cyclohexylcarbonylaminocarbonylphenyl)-N-cyclohexylcarbony-
l-2-amino ethyl]-4-(2-methoxyphenyl)piperazine
EXAMPLE 17
[0138]
1-[N-(2-methoxycarbonylphenyl)-2-aminoethyl]-4-(2-methoxyphenyl)pip-
erazine
EXAMPLE 18
[0139]
1-[N-(2-methoxycarbonylphenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-
-(2-methoxyphenyl)piperazine
EXAMPLE 19
[0140]
1-[N-(2-dimethylaminocarbonylphenyl)-2-aminoethyl]-4-(2-methoxyphen-
yl) piperazine
EXAMPLE 20
[0141] 1-[N-(2-methoxyphenyl)-2-aminoethyl]-4-(2-methoxyphenyl)
piperazine
EXAMPLE 21
[0142]
1-[N-(2-dimethylaminocarbonylphenyl)-N-cyclohexylcarbonyl-2-aminoet-
hyl]-4-(2-methoxyphenyl)piperazine
EXAMPLE 22
[0143]
1-[N-(2-trifluoromethylphenyl)-2-aminoethyl]-4-(2-methoxyphenyl)pip-
erazine
EXAMPLE 23
[0144]
1-[N-(2-methoxyphenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(2-meth-
oxy phenyl)piperazine
EXAMPLE 24
[0145]
1-[N-(2-ethylaminocarbonylphenyl)-2-aminoethyl]-4-(2-methoxyphenyl)-
piperazine
EXAMPLE 25
[0146]
1-[N-(2-ethylaminocarbonylphenyl)-N-cyclohexylcarbonyl-2-aminoethyl-
]-4-(2-methoxyphenyl)piperazine
EXAMPLE 26
[0147]
1-[N-(2-trifluoromethylphenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-
-(2-methoxyphenyl)piperazine
Preparation of N-(2-trifluoromethylphenyl cyclohexanecarboxamide
(Compound 26A)
[0148] A solution of 2-trifluoromethylaniline (3 mL), triethylamine
(3.5 mL) and CH.sub.2Cl.sub.2 (30 mL) was stirred at room
temperature under N.sub.2 and added dropwise with
cyclohexanecarbonyl chloride (3.34 mL). After 2.5 h stirring at
room temperature, the mixture was poured into H.sub.2O and
alkalinized with 1 N NaOH. The organic phase was dried on anhydrous
Na.sub.2SO.sub.4 and the crude was crystallized from EtOH to give
3.82 g (59%) of the title compound. M.p. 153-154.degree. C.
[0149] .sup.1H--NMR (CDCl.sub.3, .delta.): 8.20 (dd, 1H,
trifluoromethylphenyl ring CH), 7.60-7.40 (m, 3H,
trifluoromethylphenyl ring CHs and NH), 7.12 (ddd, 1H,
trifluoromethylphenyl ring CH), 2.30 (tt, 1H, CHC(O)), 2.10-1.20
(m, 10H, cyclohexyl protons).
Preparation of 1-[N-(2-trifluoromethylphenyl)-N-cyclohexylcarbonl
-2-aminoethyl]-4-(2-methoxyphenyl)piperazine
[0150] A mixture of
N-(2-trifluoromethylphenyl)cyclohexanecarboxamide (0.2 g) (Compd
26A), 1-(2-chloroethyl)-4-(2-methoxyphenyl)piperazine (0.37 g), 50%
(w/w) NaOH (0.5 mL), TEBAC (0.16 g) and toluene (2 mL) was stirred
at 80.degree. C. for 3.5 h. An additional amount of compd 26A (0.2
g) was then added and after 6 h stirring at 80.degree. C. the
mixture was poured into H.sub.2O and extracted with
CH.sub.2Cl.sub.2. The organic phase was dried on anhydrous
Na.sub.2SO.sub.4, evaporated to dryness and the residue purified by
flash chromatography (EtOAc-petroleum ether 3:7) giving 0.12 g
(17%) of the title compound.
[0151] .sup.1H--NMR (CDCl.sub.3, .delta.): 7.77 (dd, 1H,
trifluoromethylphenyl ring CH), 7.70-7.45 (m, 3H,
trifluoromethylphenyl ring CHs), 7.10-6.80 (m, 4H, methoxyphenyl
ring CHs), 4.70-4.50 (m, 1H, CONCH(H)CH.sub.2N), 3.85 (s, 3H,
OCH.sub.3), 3.20-2.90 (m, 5H, CONCH(H)CH.sub.2N and piperazine
protons), 2.85-2.45 (m, 7H, CHC(O), CONCH.sub.2CH.sub.2N and
piperazine protons), 1.90-0.75 (m, 10H, cyclohexyl protons).
EXAMPLE 27
[0152]
1-[N-(2-aminophenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(2-methox-
yphenyl) piperazine
EXAMPLE 28
[0153]
1-[N-(2-acetylaminophenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(2--
methoxy phenyl)piperazine
EXAMPLE 29
[0154]
1-[N-(2-nitrophenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(2-methox-
yphenyl) piperazine N.sup.1-oxide
EXAMPLE 30
[0155]
1-[N-(2-nitrophenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(2-methox-
yphenyl) piperazine N.sup.4-oxide
EXAMPLE 31
[0156]
1-[N-(2-nitrophenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(2-methox-
yphenyl) piperazine N.sup.1,N.sup.4-dioxide
EXAMPLE 32
[0157]
1-[N-(2-nitrophenyl)-N-(3-furylcarbonyl)-2-aminoethyl]-4-(2-methoxy-
phenyl) piperazine
EXAMPLE 33
[0158]
1-[N-(2-nitrophenyl)-N-(2-furylcarbonyl)-2-aminoethyl]-4-(2-methoxy-
phenyl) piperazine
EXAMPLE 34
[0159]
1-[N-(2-nitrophenyl)-N-(2-thiophenecarbonyl)-2-aminoethyl]-4-(2-met-
hoxy phenyl)piperazine
EXAMPLE 35
[0160]
1-[N-(2-nitrophenyl)-N-(3-thiophenecarbonyl)-2-aminoethyl]-4-(2-met-
hoxy phenyl)piperazine
EXAMPLE 36
[0161]
1-[N-(2-nitrophenyl)-N-(4-pyridylcarbonyl)-2-aminoethyl]-4-(2-metho-
xyphenyl) piperazine
EXAMPLE 37
[0162]
1-[N-(2-nitrophenyl)-N-(3-pyridylcarbonyl)-2-aminoethyl]-4-(2-metho-
xyphenyl) piperazine
EXAMPLE 38
[0163]
1-[N-(2-nitrophenyl)-N-(2-pyrazinylcarbonyl)-2-aminoethyl]-4-(2-met-
hoxy phenyl)piperazine
EXAMPLE 39
[0164]
1-[N-(2-nitrophenyl)-N-(1-methylcyclohexylcarbonyl)-2-aminoethyl]-4-
-(2-methoxy phenyl)piperazine
EXAMPLE 40
[0165]
1-[N-(2-nitrophenyl)-N-(1-phenylcyclohexylcarbonyl)-2-aminoethyl]-4-
-(2-methoxyphenyl)piperazine
EXAMPLE 41
[0166]
1-[N-[2-(2,2,2-trifluoroethoxy)phenyl]-2-aminoethyl]-4-(2-methoxyph-
enyl) piperazine
EXAMPLE 42
[0167]
1-[N-[2-(2,2,2-trifluoroethoxy)phenyl]-N-cyclohexylcarbonyl-2-amino-
ethyl]-4-92-methoxyphenyl)piperazine
EXAMPLE 43
[0168]
1-[N-(2-cyanophenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(2-methox-
y phenyl)piperazine hydrochloride
EXAMPLE 44
[0169]
1-[N-(2-nitrophenyl)-1-amino-2-propyl]-4-(2-methoxyphenyl)piperazin-
e
EXAMPLE 45
[0170]
1-[N-(2-nitropheny)-N-cyclohexylcarbonyl-1-amino-2-propyl]-4-(2-met-
hoxyphenyl)piperazine
EXAMPLE 46
[0171]
1-[N-cyclohexylcarbonyl-N-(2-methanesulphonylaminophenyl)-2-aminoet-
hyl]-4-(2-methoxyphenyl)piperazine
EXAMPLE 47
[0172]
1-[N-(2-methoxyphenyl)-2-aminoethyl]-4-(4-indolyl)piperazine
EXAMPLE 48
[0173]
1-[N-cyclohexylcarbonyl-N-(2-methoxyphenyl)-2-aminoethyl]-4-(4-indo-
lyl)piperazine
EXAMPLE 49
[0174]
1-[N-cyclohexylcarbonyl-N-(2-benzyloxycarbonylphenyl)-2-aminoethyl]-
-4-(2-methoxyphenyl)piperazine
EXAMPLE 50
[0175] 1-[N-(2-trifluoromethoxypheny l)-2-aminoethyl]-4-(4-indolyl)
piperazine
EXAMPLE 51
[0176]
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]--
4-(4-indolyl)piperazine
EXAMPLE 52
[0177]
1-[N-(2-nitrophenyl)-2-aminopropyl]-4-(2-methoxyphenyl)piperazine
EXAMPLE 53
[0178]
1-[N-(2-nitrophenyl)-N-(2-pyrazinecarbonyl)-2-aminoethyl)]-4-(4-ind-
olyl)piperazine
EXAMPLE 54
[0179]
1-[N-(2-trifluoromethoxyphenyl)-2-aminoethyl]-4-(2-methoxy-4-nitrop-
henyl)-piperazine
EXAMPLE 55
[0180]
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]--
4-(2-methoxy-4-nitrophenyl)piperazine
EXAMPLE 56
[0181]
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]--
4-(4-amino-2-methoxyphenyl)piperazine
EXAMPLE 57
[0182]
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]--
4-(4-acetylamino-2-methoxyphenyl)piperazine
EXAMPLE 58
[0183]
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]--
4-(4-trifluoroacetylamino-2-methoxyphenyl)piperazine
EXAMPLE 59
[0184]
1-[N-cyclohexylcarbonyl-N-(2-propoxycarbonylphenyl)-2-aminoethyl]-4-
-(2-methoxyphenyl)piperazine
EXAMPLE 60
[0185]
1-[N-cyclohexylcarbonyl-N-(2-nitrophenyl)-2-aminoethyl]-4-[4-(N-ace-
tyl-N-cyclohexylcarbonyl)amino-2-methoxyphenyl]piperazine
EXAMPLE 61
[0186]
1-[N-cyclohexylcarbonyl-N-(2-nitrophenyl)-2-aminoethyl]-4-[4-(bis-c-
yclohexylcarbonyl)amino-2-methoxyphenyl]piperazine
EXAMPLE 62
[0187]
1-[N-cyclohexylcarbonyl-N-(2-nitrophenyl)-2-aminoethyl]-4-[4-cycloh-
exanecarbonylamino-2-methoxyphenyl)piperazine
EXAMPLE 63
[0188]
1-[N-(2-nitrophenyl)-N-(2-pyrimidinecarbonyl)-2-aminoethyl)]-4-(2-m-
ethoxyphenyl)piperazine
EXAMPLE 64
[0189]
1-[N-[2-(2,2,2-trifluoroethoxy)phenyl)]-2-aminoethyl]-4-(2-methoxy--
4-nitrophenyl)piperazine
EXAMPLE 65
[0190]
1-[N-cyclohexylcarbonyl-N-[2-(2,2,2-trifluoroethoxyphenyl)]-2-amino-
ethyl)]-4-(2-methoxy-4-nitrophenyl)piperazine
EXAMPLE 66
[0191]
1-[N-cyclohexylcarbonyl-N-[2-(2,2,2-trifluoroethoxyphenyl)]-2-amino-
ethyl)]-4-(4-amino-2-methoxyphenyl)piperazine
EXAMPLE 67
[0192]
1-[N-cyclohexylcarbonyl-N-[2-(2,2,2-trifluoroethoxyphenyl)]-2-amino-
ethyl)]-4-(4-acetylamino-2-methoxyphenyl)piperazine
EXAMPLE 68
[0193]
1-[N-[2-(2,2,2-trifluoroethoxy)phenyl)]-2-aminoethyl]-4-(4-indolyl)-
piperazine
EXAMPLE 69
[0194]
1-[N-cyclohexylcarbonyl-N-[2-(2,2,2-trifluoroethoxyphenyl)]-2-amino-
ethyl]-4-(4-indolyl)piperazine
EXAMPLE 70
[0195]
1-[N-cyclohexylcarbonyl-N-(2-nitrophenyl)-2-aminoethyl)]-4-(4-acety-
lamino-2-methoxyphenyl)piperazine
EXAMPLE 71
[0196]
1-[N-(2-trifluoromethoxyphenyl)-2-aminoethyl]-4-(2-trifluoromethoxy
phenyl)piperazine
EXAMPLE 72
[0197]
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]--
4-(2-trifluoromethoxyphenyl)piperazine
EXAMPLE 73
[0198]
1-[N-(2-nitrophenyl)-N-(5-thiazolylcarbonyl)-2-aminoethyl)]-4-(2-me-
thoxyphenyl)piperazine
EXAMPLE 74
[0199]
1-[N-(2-trifluoromethoxyphenyl)-2-aminoethyl]-4-(2-bromo-5-methoxyb-
enzyl)piperazine
EXAMPLE 75
[0200]
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]--
4-(2-bromo-5-methoxybenzyl)piperazine
EXAMPLE 76
[0201]
1-[N-cyclohexylcarbonyl-N-(2-iodophenyl)-2-aminoethyl]-4-(4-indolyl-
)piperazine
EXAMPLE 77
[0202]
1-[N-(2-cyanophenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(2-triflu-
oromethoxyphenyl)piperazine
EXAMPLE 78
[0203]
1-[N-(2-cyanophenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(2-methox-
y-4-nitrophenyl)piperazine
EXAMPLE 79
[0204]
1-[N-(2-cyanophenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(4-amino--
2-methoxyphenyl)piperazine
EXAMPLE 80
[0205]
1-[N-(2-cyanophenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(4-acetyl-
amino-2-methoxyphenyl)piperazine
EXAMPLE 81
[0206]
1-[N-(2-acetylphenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(2-metho-
xyphenyl)piperazine
EXAMPLE 82
[0207]
1-[N-(2-cyanophenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(4-indoly-
l)piperazine
EXAMPLE 83
[0208]
1-[N-(2-cyanophenyl)-N-cyclohexylcarbonyl-2-aminoethyl]-4-(2-bromo--
5-methoxybenzyl)piperazine
EXAMPLE 84
[0209]
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]--
4-(1 -cyclohexylcarbonyl-4-indolyl)piperazine
EXAMPLE 85
[0210]
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]--
4-(8-quinolyl)piperazine
Preparation of N-(2-trifluoromethoxyphenyl)cyclohexanecarboxamide
(Compound 85A)
[0211] This compound was prepared following the procedure described
for compound 26A of example 26, except that
2-trifluoromethoxyaniline was used in place of
2-trifluoromethylaniline. Yield: 100%.
[0212] .sup.1H--NMR (CDCl.sub.3, .delta.): 8.44 (dd, 1H, phenyl
ring H6), 7.47 (bs, 1H, CONH), 7.21-730 (m, 2H, phenyl ring H4,
H5), 7.08-7.14 (m, 1H, phenyl ring H3), 2.21-2.43 (m, 1H, CHCO),
1.20-2.08 (m, 10H, cyclohexyl protons).
Preparation of N-(2
,2-dimethoxyethyl)-N-(2-trifluoromethoxyphenyl)cyclohe-
xanecarbox-amide (85B)
[0213] A solution of compound 85A (0.15 g) in toluene (20 mL) was
heated to reflux and 5 mL of toluene distilled off; then t-BuOK
(0.09 g) was added and the mixture heated to reflux removing by
evaporation 2 mL of the solvent; then 2-bromoacetaldehyde dimethyl
acetal (0.09 mL) was dropped. The resulting suspension was refluxed
under N.sub.2 atmosphere for 12-15 h. After cooling to room
temperature, the solvent was evaporated to dryness in vacuo and the
crude purified by flash chromatography (petroleum ether--EtOAc
9:1), affording 0.106 g (54%) of the title compound as an oil.
[0214] .sup.1H--NMR (CDCl.sub.3, .delta.): 7.31-7.48 (m, 4H, phenyl
ring CHs), 4.61-4.70 (m, 1H, CH(OCH.sub.3).sub.2), 4.16-4.24 (m,
1H, CONCH(H)), 3.39 (s, 3H, OCH.sub.3),3.32 (s, 3H, OCH.sub.3),
3.13-3.24 (m, 1H, CONCH(H)), 1.92-2.09 (m, 1H, CHCO), 0.85-1.79 (m,
10H, cyclohexyl protons).
Preparation of
N-(2-oxoethyl)-N-(2-trifluoromethoxyphenyl)cyclohexanecarbo- xamide
(85C)
[0215] A suspension of compound 85B (0.106 g), hydroquinone (0.003
g) in 2 N HCl (1.14 mL) was heated at 80.degree. C. for 30-40
minutes. After cooling to room temperature, NaHCO.sub.3 was added
(pH=7) and the resulting mixture was extracted with
CH.sub.2Cl.sub.2 (3.times.10 mL).The combined organic layers were
dried (Na.sub.2SO.sub.4), filtered and evaporated to dryness at
reduced pressure to give 0.081 g (88%) of the title compound as an
oil.
[0216] .sup.1H--NMR (CDCl.sub.3, .delta.): 9.62 (s, 1H, CHO),
7.29-7.57 (m, 4H, phenyl ring CHs), 4.67 (d, 1H, NCH(H)CHO), 3.86
(d, 1H, NCH(H)CHO), 2.02-1.98 (m, 1H, CHCO), 0.85-1.83 (m, 10H,
cyclohexyl protons).
Preparation of 1-(8-quinolyl]piperazine (85D)
[0217] A mixture of 8-aminoquinoline (1.5 g),
bis-(2-chloroethyl)amine hydrochloride (2.04 g), o-dichlorobenzene
(4.5 mL) and n-hexanol (0.45 mL) was stirred at reflux temperature
for 5 h. After cooling to room temperature, the mixture was treated
with 2 N NaOH (10 mL) and extracted with CH.sub.2Cl.sub.2
(3.times.20 mL). The purification was carried out by flash
chromatography (CH.sub.2Cl.sub.2-2 N methanolic NH.sub.3) 97:3 to
give 0.48 g (22%) of the title compound.
[0218] .sup.1H--NMR (CDCl.sub.3, .delta.): 8.80 (dd, 1H, quinolyl
H2), 8.17 (dd, 1H, quinolyl H4), 7.32-7.53 (m, 3H, quinolyl H6, H5,
H3), 7.12 (m, 1H, quinolyl H7), 3.38-3.51 (m, 4H, piperazine
protons), 3.21-3.32 (m, 4H, piperazine protons), 2.21 (bs, 1H,
NH).
Preparation of
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-amin-
oethyl]-4-(8-quinolyl)piperazine
[0219] To a solution of compound 85C (0.081 g), compound 85D (0.06
g) and acetic acid (0.06 mL) in 1,2-DCE (10 mL) stirred under
N.sub.2 atmosphere, NaB(Oac).sub.3H (0.078 g) was added. The
resulting mixture was stirred for 2 h at 20-25.degree. C.,
alkalinized with 1 N NaOH (pH=8) and extracted with
CH.sub.2Cl.sub.2 (3.times.10 mL). The organic layer was dried over
Na.sub.2SO.sub.4, filtered and evaporated at reduced pressure. The
crude was purified by flash chromatography (toluene-acetone 75:25)
to give 0.09 g (73%) of the title compound. M.p. 141.5-143.degree.
C.
[0220] .sup.1H--NMR (CDCl.sub.3, .delta.): 8.86 (dd, 1H, quinolyl
H2), 8.14 (dd, 1H, quinol H4), 7.30-7.52 (m, 7H, quinolyl H3, H5,
H6 and phenyl ring CHs), 7.09-7.18 (m, 1H, quinolyl H7), 4.32-4.47
(m, 1H, CONCH(H)CH.sub.2N), 3.21-3.51 (m, 5H, piperazine protons
and CONCH(H)CH.sub.2N), 2.53-2.94 (m, 6H, piperazine protons and
CONCH.sub.2CH.sub.2N), 1.89-2.07 (m, 1H, CHCO), 0.81-1.79 (m, 10H,
cyclohexyl protons).
EXAMPLE 86
[0221] 1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl
)-2-aminoethyl]-4-(4-amino-2-cyanophenyl)piperazine
EXAMPLE 87
[0222] 1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl
)-2-aminoethyl]-4-(2-cyanophenyl)piperazine
EXAMPLE 88
[0223]
1-[N-cyclohexylcarbonyl-N-(2-methanesulphonylaminophenyl)-2-aminoet-
hyl]-4-(4-indolyl)piperazine
EXAMPLE 89
[0224]
1-[N-cyclohexylcarbonyl-N-(2-methoxyphenyl)-2-aminoethyl]-4-(1-cycl-
ohexylcarbonyl-4-indolyl)piperazine
EXAMPLE 90
[0225]
1-[N-cyclohexylcarbonyl-N-(2-methoxyphenyl)-2-aminoethyl]-4-[4-fluo-
ro-2-(2,2,2-trifluoroethoxy)phenyl]piperazine
Preparation of [2-(2-methoxyphenyl)amino]acetaldehyde dimethyl
acetal (Compound 90A)
[0226] A stirred mixture of 5.8 mL of 2-methoxyaniline, 50 mL of
anhydrous DMF, 20.75 g of anhydrous K.sub.2CO.sub.3 and 15.5 mL of
97% 2-bromoacetaldehyde dimethyl acetal was heated at 150.degree.
C. for 5 h. After diluting with H.sub.2O (500 mL), the mixture was
extracted with Et.sub.2O (4.times.70 mL); the organic layer was
washed with H.sub.2O (5.times.50 mL), dried (anhydrous
Na.sub.2SO.sub.4) and evaporated to dryness in vacuo. The residue
was purified by flash chromatography (petroleum ether-EtOAc 90:10)
to afford 9.5 g (90.5%) of Compond 90A as a yellow oil.
[0227] .sup.1H--NMR (CDCl.sub.3, .delta.): 3.26 (d, 2H, CH.sub.2),
3.43 (s, 6H, 2 CH.sub.3O), 3.85 (s, 3H, CH.sub.3O), 4.00-4.70 (br,
1H, NH), 4.61 (t, 1H, CH), 6.56-6.95 (m, 4H, phenyl CHs) After
D.sub.2O treatment the NH signal appeared as HDO peak.
Preparation of N-(2
,2-dimethoxyethyl)-N-(2-methoxyphenyl)cyclohexanecarbo- xamide
(Compound 90B)
[0228] A stirred mixture of 0.73 g of Compound 90A, 40 mL of
CH.sub.2Cl.sub.2, 1 mL of 97% Et.sub.3N and 0.60 mL (4.4 mmol) of
98% cyclohexanecarbonyl chloride was maintained at room temperature
for 24 h, washed with H.sub.2O (5.times.20 mL), 2 N NaOH
(6.times.20 mL), dried (anhydrous Na.sub.2SO.sub.4) and evaporated
to dryness in vacuo. The residue was purified by flash
chromatography (petroleum ether-EtOAc 80:20) to afford 0.955 g
(86%) of Compound 90B as an oil.
[0229] .sup.1H--NMR (CDCl.sub.3,.delta.): 0.75-1.75 (m, 10H,
cyclohexane CH.sub.2s), 1.90-2.15 (m, 1H, CHCO), 3.18 (dd, 1H,
CONCH(H)CH), 3.25 and 3.35 (2s, 6H, 2 CH.sub.3O), 3.81 (s,3H,
CH.sub.3O), 4.20 (dd, 1H, CONCH(H)CH), 4.60(ddd, 1H, CONCH(H)CH),
6.85-7.05 (m, 2H, phenyl CHs), 7.15-7.40 (m, 2H, phenyl CHs)
Preparation of
N-(2-oxoethyl)-N-(2-methoxyphenyl)cyclohexanecarboxamide (Compound
90C)
[0230] A stirred mixture of 0.955 g of Compound 90B, 0.10 g of
hydroquinone and 20 mL of 2 N HCl was heated to 80.degree. C. for
30' under nitrogen atmosphere. After cooling at 0-5.degree. C., 60
mL of CH.sub.2Cl.sub.2 was added and the mixture was alkalinized
with 20% aq. Na2CO.sub.3. The layers were separated and the
alkaline one was re-extracted with CH.sub.2Cl.sub.2 (2.times.40
mL). The combined organic layers were dried (anhydrous
Na.sub.2SO.sub.4) and evaporated to dryness in vacuo to afford
0.646 g (79%) of Compound 90C as an orange oil.
[0231] .sup.1H--NMR (CDCl.sub.3, .delta.): 0.80-1.85 (m, 10H,
cyclohexane CH.sub.2s), 2.10-2.30 (m, 1H, CHCO), 3.82 (s, 3H,
CH.sub.3O), 4.18 (d, 2H, CH.sub.2), 6.90-7.05 (m, 2H, phenyl CHs),
7.20-7.45(m, 2H, phenyl CHs), 9.65 (s, 1H, CHO)
Preparation of
1-[N-cyclohexylcarbonl-N-(2-methoxyphenyl)-2-aminoethyl]-4--
[4-fluoro-2-(2,2 ,2-trifluoroethoxy)phenyl]piperazine
[0232] The title compound was prepared following the method
described in example 85 substituting compound 90C for compound 85C
and 1-[4-fluoro-2-(2,2,2-trifluoroethoxy)phenyl]piperazine for
1-(8-quinolyl)piperazine. The residue was purified by flash
chromatography (CHCl.sub.3-2 N methanolic ammonia 100:1) to afford
the title compound g (57%) as a yellow oil.
[0233] .sup.1H--NMR (CDCl.sub.3, .delta.): 0.80-1.75 (m, 1OH,
cyclohexane CH.sub.2s), 1.90-2.12 (m, 1H, CHCO), 2.40-2.75 (m, 6H,
CONCH(H)CH.sub.2, piperazine CHs), 2.85-3.10 (m, 4H, piperazine
CHs), 3.30-3.50 (m, 1H, CONCH(H)CH.sub.2), 3.82 (s, 3H, CH.sub.3O),
4.08-4.28 (m, 1H,CONCH(H) CH.sub.2), 4.38 (q, 2H,
OCH.sub.2CF.sub.3), 6.58-6.80(m, 2H, fluorophenyl H3 and H6),
6.80-7.05 (m, 3H, fluorophenyl H5, methoxyphenyl CHs), 7.15-7.43
(m, 2H, methoxyphenyl CHs)
EXAMPLE 91
[0234]
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]--
4-[4-fluoro-2-(2,2,2-trifluoroethoxy)phenyl]piperazine
[0235] The title compound was prepared following the method
described in example 85 substituting
1-[4-fluoro-2-(2,2,2-trifluoroethoxy)phenyl]piper- azine for
1-(8-quinolyl)piperzine. The residue was purified by flash
chromatography (CHCl.sub.3-2 N methanolic ammonia 100:1) to afford
the title compound (90%) as a yellow oil.
[0236] .sup.1H--NMR (CDCl.sub.3, .delta.): 0.75-1.78 (m, 10H,
cyclohexane CH.sub.2s), 1.80-2.08 (m, 1H, CHCO), 2.40-2.75 (m, 6H,
CONCH(H)CH.sub.2, piperazine CHs), 2.85-3.10 (m, 4H, piperazine
CHs), 3.18-3.40 (m, 1H, CONCH(H)CH.sub.2), 4.20-4.48 (m, 3H,
CF.sub.3CH.sub.2O, CONCH(H)CH.sub.2), 6.58-6.80 (m, 2H,
fluorophenyl H3 and H6), 6.80-6.97 (m, 1H, fluorophenyl H5),
7.21-7.50 (m, 4H, trifluoromethoxyphenyl CHs)
EXAMPLE 92
[0237]
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]--
4-(2,4-dimethoxyphenyl)piperazine
[0238] The title compound was prepared following the method
described in example 85 substituting
1-(2,4-dimethoxyphenyl)piperazine for 1-(8-quinolyl)piperazine. The
residue was purified by flash chromatography (CH.sub.2Cl.sub.2-2 N
methanolic ammonia 100:1) to afford the title compound (80%) as an
ivory oil.
[0239] .sup.1H--NMR (CDCl.sub.3, .delta.): 0.85-1.78 (m, 10H,
cyclohexane CH.sub.2s), 1.78-2.05 (m, 1H, CHCO), 2.40-2.75 (m, 6H,
CONCH(H)CH.sub.2, piperazine CHs), 2.80-3.05 (m, 4H, piperazine
CHs), 3.15-3.40 (m, 1H, CONCH(H)CH.sub.2), 3.75 and 3.80 (s, 6H, 2
CH.sub.3O), 4.20-4.40 (m, 1H, CONCH(H)CH.sub.2), 6.32-6.50 (m, 2H,
methoxyphenyl H3 and H5), 6.80 (d, 1H, methoxyphenyl H6), 7.20-7.50
(m, 4H, trifluoromethoxyphenyl CHs)
EXAMPLE 93
[0240]
1[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]-4-
-(4-hydroxy-2-methoxyphenyl)piperazine
[0241] The title compound was prepared following the method
described in example 85 substituting
1-(4-hydroxy-2-methoxyphenyl)piperazine.2H.sub.2O for
1-(8-quinolyl)piperazine. The residue was purified by flash
chromatography (CHCl.sub.3-2 N methanolic ammonia 100:2) to afford
the title compound (55%) as an ivory amorphous solid.
[0242] .sup.1H--NMR (CDCl.sub.3, .delta.): 0.75-1.75 (m, 10H,
cyclohexane CH.sub.2s), 1.82-2.08 (m, 1H, CHCO), 2.40-2.75 (m, 6H,
CONCH(H)CH.sub.2, piperazine CHs), 2.75-3.05 (m, 4H, piperazine
CHs), 3.15-3.40 (m, 1H, CONCH(H)CH.sub.2), 3.80 (s, 6H, CH.sub.3O),
4.20-4.40 (m, 1H, CONCH(H)CH.sub.2), 4.65-5.50 (br, 1H, OH), 6.32
(dd, 1H, methoxyphenyl H5), 6.42 (d, 1H, methoxyphenyl H3), 6.78
(d, 1H, methoxyphenyl H6), 7.20-7.50 (m, 4H,
trifluoromethoxy-phenyl CHs)
EXAMPLE 94
[0243]
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]--
4-(4-acetoxy-2-methoxyphenyl)piperazine
[0244] To a mixture of 0.15 g of the compound of Example 93, 20 mL
of CH.sub.2Cl.sub.2 and 0.06 mL of 97% Et.sub.3N was added 0.03 mL
of 98% CH.sub.3COCl at r.t. After 6 h at r.t. the reaction solution
was diluted with CH.sub.2Cl.sub.2 (20 mL), washed with H.sub.2O
(5.times.10 mL), 1 N NaOH (2.times.10 mL), H.sub.2O (2.times.10
mL), dried (anhydrous Na.sub.2SO.sub.4) and evaporated to dryness
in vacuo. The residue was purified by flash chromatography
(CH.sub.2Cl.sub.2 -2 N methanolic ammonia 100:3) to afford 0.12 g
(73.7%) of the title compound as a yellow oil.
[0245] .sup.1H--NMR (CDCl.sub.3, .delta.): 0.85-1.80 (m, 10H,
cyclohexane CH.sub.2s), 1.80-2.05 (m, 1H, CHCO), 2.28 (s, 3H,
CH.sub.3COO), 2.40-2.80 (m, 6H, CONCH(H)CH.sub.2, piperazine CHs),
2.80-3.15 (m, 4H, piperazine CHs), 3.15-3.40 (m, 1H,
CONCH(H)CH.sub.2), 3.80 (s, 3H, CH.sub.3O), 4.10-4.60 (m, 1H,
CONCH(H)CH.sub.2), 6.50-6.70 (m, 2H, methoxyphenyl H3 and H5), 6.85
(d, 1H, methoxyphenyl H6), 7.20-7.50 (m, 4H, trifluoromethoxyphenyl
CHs)
EXAMPLE 95
[0246]
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]--
4-(4-ethylaminocarbonyloxy-2-methoxyphenyl)piperazine
[0247] To a stirred solution of 0.24 g (0.46 mmol) of the compound
of example 93 in 20 mL of anhydrous THF was added 0.02 g (0.50
mmol) of 60% NaH at r.t. under nitrogen. After stirring for 1 h at
r.t., 0.05 mL (0.62 mmol) of 98% ethyl isocyanate was added and the
mixture was stirred for 4 h, diluted with H.sub.2O (50 mL) and
brine (10 mL) and extracted with EtOAc (3.times.30 mL). The organic
layer was dried (anhydrous Na.sub.2SO.sub.4) and evaporated to
dryness in vacuo. The residue was purified by flash chromatography
(CHCl.sub.2-2 N methanolic ammonia 100:2) to afford 0.15 g (54.4%)
of the title compound as a vitreous ivory solid.
[0248] .sup.1H--NMR (CDCl.sub.3, .delta.): 0.75-1.80 (m, 13H,
cyclohexane CH.sub.2s, CH.sub.3CH.sub.2N), 1.80-2.08 (m, 1H, CHCO),
2.40-2.80 (m, 6H, CONCH(H)CH.sub.2, piperazine CHs), 2.80-3.15 (m,
4H, piperazine CHs), 3.15-3.45 (m, 1H, CONCH(H)CH.sub.2), 3.80 (s,
3H, CH.sub.3O), 3.90-4.10 (m, 1H, CH.sub.3CH.sub.2N), 4.20-4.40 (m,
1H, CONCH(H)CH.sub.2), 6.50-6.75 (m, 2H, methoxyphenyl H3 and H5),
6.90 (d, 1H, methoxyphenyl H6), 7.20-7.50 (m, 4H,
trifluoromethoxyphenyl CHs), 8.20-8.58 (m, 1H, CONH)
EXAMPLE 96
[0249]
1-[N-cyclohexylcarbonyl-N-(2-trifluoromethoxyphenyl)-2-aminoethyl]--
4-[4-(2-methylpropionyloxy)-2-methoxyphenyl]piperazine
[0250] The title compound was prepared following the procedure
described for compound 94 but using 2-methylpropionyl chloride
instead of acetyl chloride. The residue was purified by flash
chromatography (EtOAc-petroleum ether 1:1) to afford the title
compound (17%) as a yellow oil.
[0251] .sup.1H--NMR (CDCl.sub.3, .delta.): 0.75-1.75 (m, 16H,
cyclohexane CH.sub.2s, CH(CH.sub.3).sub.2), 1.80-2.05 (m, 1H,
CHCO), 2.35-2.85 (m, 7H, CONCH(H)CH.sub.2, piperazine CHs,
CH(CH.sub.3).sub.2), 2.85-3.10 (m, 4H, piperazine CHs), 3.15-3.40
(m, 1H, CONCH(H)CH.sub.2), 3.80 (s, 3H, CH.sub.3O), 4.20-4.40 (m,
1H, CONCH(H)CH.sub.2), 6.50-6.70 (m, 2H, methoxyphenyl H3 and H5),
6.85 (d, 1H, methoxyphenyl H6), 7.20-7.50 (m, 4H,
trifluoromethoxyphenyl CHs).
EXAMPLE 97
Effects on Volume-Induced Rhythmic Bladder Voiding Contractions in
Anaesthetized Rats
A. Methods
[0252] Female Sprague Dawley rats weighing 225-275 g (Crl: CDo BR,
Charles River Italia) were used. The animals were housed with free
access to food and water and were maintained on a forced 12 h
alternating light-dark cycle at 22-24.degree. C. for at least one
week, except during the experiment. The activity on the rhythmic
bladder voiding contractions was evaluated according to the method
of Dray (J. Pharmacol. Methods, 13:157, 1985), with some
modifications as in Guarneri (Pharmacol. Res., 27:173, 1993).
Briefly, rats were anesthetized by subcutaneous injection of 1.25
g/kg (5 ml/kg) urethane, after which the urinary bladder was
catheterized via the urethra using PE 50 polyethylene tubing filled
with physiological saline. The catheter was tied in place with a
ligature around the external urethral orifice and was connected
with conventional pressure transducers (Statham P23 ID/P23 XL). The
intravesical pressure was displayed continuously on a chart
recorder (Battaglia Rangoni KV 135 with DCl/TI amplifier). The
bladder was then filled via the recording catheter by incremental
volumes of warm (37.degree. C.) saline until reflex bladder voiding
contractions occurred (usually 0.8-1.5 ml). For intravenous (i.v.)
injection of bioactive compounds, PE 50 polyethylene tubing filled
with physiological saline was inserted into the jugular vein.
[0253] From the cystometrogram, the number of contractions recorded
15 min before (basal values) and after treatment, as well as the
mean amplitude of these contractions (mean height of the peaks in
mmHg) was evaluated. Since most compounds produced an effect that
was relatively rapid in onset and led to a complete cessation of
bladder contractions, bioactivity was conveniently estimated by
measuring the duration of bladder quiescence (i.e., the duration of
time during which no contractions occurred). The number of animals
tested showing a reduction in the number of contractions >30% of
that observed in the basal period was also recorded.
[0254] To compare the potency of the tested compounds for
inhibiting bladder voiding contractions, equieffective doses which
resulted in a contraction disappearance time of 10 minutes
(ED.sub.10min) were computed by means of least square linear
regression analysis. Also computed in this manner were extrapolated
doses which induced a reduction of the number of contractions of
greater than 30% in 50% of treated rats (ED.sub.50, frequency) by
the method of Bliss (Bliss C. I., Quart. J. Pharm. Pharmacol. 11,
192-216, 1938). After the suppressive effects of drug injection
wore off, the height of the contractile peaks was compared with the
height of the peaks previously recorded after the control
intravenous administration of vehicle. The potency of the tested
compounds (ED.sub.50 value: the extrapolated doses inducing a 30%
reduction of amplitude of the contractions in 50% of treated rats)
was evaluated on a quantal basis by the method of Bliss (Bliss C.
I., Quart. J. Pharm. Pharmacol. 11, 192-216, 1938).
B. Results
[0255] The rapid distension of the urinary bladder in
urethane-anesthetized rats produced a series of rhythmic bladder
voiding contractions whose characteristics have been described and
are well-known in the art (Maggi et al., Brain Res., 380:83, 1986;
Maggi, et al., J. Pharmacol. Exp. Ther., 230:500, 1984). The
frequency of these contractions is related to the sensory afferent
arm of reflex micturition and to the integrity of the micturition
center, while their amplitude is a property of the efferent arm of
the reflex. In this model system, compounds that act mainly on the
CNS (such as morphine) cause a block in voiding contraction,
whereas drugs that act at the level of the detrusor muscle, such as
oxybutynin, lower the amplitude of the bladder contractions.
[0256] The results obtained after administration of prior art
compounds and compounds of the invention are shown in Table 1.
Compound A, a prior art compound, was more potent than flavoxate
and oxybutynin in inhibiting voiding contractions. This compound,
in contrast to oxybutynin, did not affect the amplitude of the
contraction, indicating no impairment of bladder contractility.
Surprisingly, however, compounds with substituents at position 2 of
the aniline ring in Formula I, such as NO.sub.2, such as the
compound of Example 2, have significantly higher potency than
unsubstituted compound A, particular with regard to the
ED.sub.10min values. Like compound A, the compound of Example 2
does not affect bladder contractility. When compounds were
synthesized with a nitro group at position 3 or 4 of the aniline
ring, such as comparative compounds B and C, pharmacological
activity was abolished.
[0257] Results similar (i.e., higher potency for the 2-substituted
derivatives) to those obtained by Example 2 were obtained relative
to compounds where R is H and which are unsubstituted or
substituted at the 3- and 4-positions of the aniline ring. These
results are shown in Table 1, where it can be seen that compounds
AA, D, E, which are unsubstituted and 3- and 4-substituted
compounds, are clearly inferior to the compounds of Examples 1, 10
and 11 and 18, i.e., 2-NO.sub.2, 2-CN, 2-COCH.sub.3 and
2-COOCH.sub.3 derivatives. The compounds of the invention were
clearly superior, particularly with regard to the ED.sub.50 values
which are indicators of urinary frequency.
1TABLE 1 Effects on rhythmic bladder voiding contractions after
intravenous administration. Data represent the ED.sub.10 min values
(the extrapolated dose inducing 10 min of disappearance of the
contractions); the ED.sub.50 values (the extrapolated doses
inducing a reduction of the number of contractions >30% in 50%
of treated rats) (frequency), and the ED.sub.50 values (the
extrapolated doses inducing 30% reduction of amplitude of the
contractions in 50% of treated rats) (amplitude). ED50 ED50
Compound ED.sub.10 min .mu.g/kg (frequency) .mu.g/kg (amplitude)
.mu.g/kg Compound A 650 33 n.a. Compound B >1000 >1000 n.a.
Compound C >1000 >1000 n.a. Compound D >1000 >1000 n.a.
Compound E >1000 >1000 n.a. Compound AA 663 244 n.a. Example
1 192 55 n.a. Example 2 60 9 n.a. Example 10 122 28 n.a. Example 11
318 40 n.a. Example 13 266 29 n.a. Example 18 101 17 n.a. Example
20 97 25 n.a. Example 23 93 18 n.a. Example 27 131 13 n.a. Example
42 33 18 n.a. Example 43 52 4 n.a. Example 47 181 20 n.a. Example
48 40 14 n.a. Example 51 276 200 n.a. Example 56 61 21 n.a. Example
57 254 252 n.a. Example 66 167 58 n.a. Example 69 60 14 n.a.
Flavoxate >10000 2648 n.a. Oxybutinin 7770 >10000 240 n.a. =
not active; no significant reduction of the height of peaks.
Compound AA = 1-(N-phenyl-2-aminoethyl)-4-(2-methoxyphenyl-
)piperazine. Compounds A, B, C, D, E, see text.
EXAMPLE 98
Effects on Cystometric Parameters in Conscious Rats
A. Methods
[0258] Male Sprague Dawley rats (Crl: CDo BR) weighing 250-350 g
were used. The animals were housed with free access to food and
water and maintained on a forced 12 h alternating light-dark cycle
at 22-24.degree. C. for at least one week, except during
performance of the experiment. To quantify urodynamic parameters in
conscious rats, cystometrographic studies were performed using
procedures previously described (Guarneri et al., Pharmacol. Res.,
24:175, 1991). Male rats were anesthetized with nembutal (30 mg/kg)
and chloral hydrate (125 mg/kg) i.p. and were placed in a supine
position. An approximately 10 mm long midline incision was made in
the shaved and cleaned abdominal wall. The urinary bladder was
gently freed from adhering tissues, emptied, and then cannulated,
via an incision at the dome, with a polyethylene cannula (Portex
PP30), which was permanently sutured with silk thread. The cannula
was exteriorized through a subcutaneous tunnel in the retroscapular
area, where it was connected with a plastic adapter to avoid the
risk of removal by the animal. For intravenous (i.v.) injection of
test compounds, a PE 50 polyethylene tubing filled with
physiological saline was inserted into the jugular vein and
exteriorized in the retroscapular area. The rats were utilized
exclusively one day after implantation. On the day of the
experiment, the rats were placed in Bollman's cages; after a
stabilization period of 20 min, and the free tip of the bladder
catheter was connected through a T-shaped tube to a pressure
transducer (Bentley T 800/Marb P 82) and to a peristaltic pump
(Gilson minipuls 2) for a continuous infusion, at the constant rate
of 0.1 ml/mmin, of saline solution into the urinary bladder. The
intraluminal pressure signal during infusion was continuously
recorded on a polygraph (Battaglia Rangoni KO 380 with ADCl/T
amplifier). Two urodynamic parameters were evaluated: bladder
volume capacity (BVC) and micturition pressure (MP). BVC (in ml) is
defined as the minimum volume infused after which detrusor
contraction (followed by micturition) occurs. MP (in mm Hg) is
defined as the maximal intravesical pressure induced by the
contraction of detrusor during micturition. Basal BVC and MP values
were calculated as the means of the first two recorded
cystometrograms. At this point in the assay, the infusion was
interrupted and the test compounds were administered. Fifteen
minutes after intravenous administration two additional
cystometrograms were recorded in each animal and the mean values of
the two cystometrographic parameters were calculated. The
statistical significance of the differences in urodynamic parameter
values was evaluated by Student's t test for paired data.
B. Results
[0259] The effects of different doses of the tested compounds are
shown in Table 2. Compound A behaved similarly to flavoxate by
increasing BVC. Neither compound impaired bladder contractility,
since no consistent changes in MP were observed. In contrast,
oxybutynin markedly and dose-dependently decreased MP without
effects on BVC. The compound of Example 2 was more potent than
compound A and flavoxate; a significant increase in BVC was
observed after the i.v. administration of 0.3 mg/kg of the compound
of Example 2, compared with the requirement for administration of
1.0 mg/kg of flavoxate or compound A. The compound of Example 2
induced a slight, albeit significant, decrease in MP. This effect,
however, was not dose-dependent and was markedly lower than that
induced by oxybutynin.
2TABLE 2 Effects on cystometrogram in conscious rats. Data
represent mean values .+-. S.E. of bladder volume capacity (BVC;
ml) and of micturition pressure (MP; mmHg), before and 15 min after
i.v. injection of the compounds. Dose BVC % of COMPOUND .mu.g/kg
before after treatment change Compound A 300 0.81 .+-. 0.05 0.87
.+-. 0.05 +7.4 1000 0.78 .+-. 0.11 0.97 .+-. 0.11** +24.4 Example 2
300 0.71 .+-. 0.09 0.87 .+-. 0.10* +22.5 1000 0.62 .+-. 0.09 0.75
.+-. 0.10** +21.0 Example 8 300 0.59 .+-. 0.04 0.71 .+-. 0.05*
+21.0 1000 0.65 .+-. 0.10 0.88 .+-. 0.12** +35.0 Example 13 100
0.94 .+-. 0.12 1.07 .+-. 0.14 +13.4 300 0.73 .+-. 0.09 0.95 .+-.
0.12** +30.2 Example 18 100 0.60 .+-. 0.07 0.80 .+-. 0.09* +33.3
1000 0.63 .+-. 0.11 0.89 .+-. 0.16** +40.3 Example 23 300 0.50 .+-.
0.06 0.77 .+-. 0.03** +54.3 1000 0.66 .+-. 0.09 0.89 .+-. 0.12**
+34.5 Example 42 300 0.70 .+-. 0.11 0.89 .+-. 0.15 +26.4 1000 0.70
.+-. 0.09 1.00 .+-. 0.16* +41.7 Example 51 300 0.66 .+-. 0.11 0.84
.+-. 0.14** +27.7 1000 0.79 .+-. 0.05 1.08 .+-. 0.09** +36.5
FLAVOXATE 300 0.76 .+-. 0.11 0.87 .+-. 0.11 +14.5 1000 0.88 .+-.
0.15 1.11 .+-. 0.16** +26.1 OXYBUTYNIN 100 0.82 .+-. 0.15 0.89 .+-.
0.18 +8.5 300 0.83 .+-. 0.13 0.83 .+-. 0.12 .+-.0.0 1000 0.94 .+-.
0.19 1.00 .+-. 0.18 .+-.6.4 Dose MP % of COMPOUND .mu.g/kg before
after treatment change Compound A 300 90.6 .+-. 10.4 85.6 .+-. 11.3
-5.5 1000 90.2 .+-. 6.5 84.1 .+-. 5.2 -6.8 Example 2 300 95.4 .+-.
6.4 80.4 .+-. 6.5** -15.7 1000 109.0 .+-. 12.1 99.6 .+-. 11.2* -8.6
Example 8 300 116.1 .+-. 17.4 98.3 .+-. 17.2** -15.0 1000 81.3 .+-.
9.0 64.8 .+-. 10.5* -20.0 Example 13 100 85.7 .+-. 14.2 75.6 .+-.
13.7 -12.5 300 73.4 .+-. 11.8 65.7 .+-. 13.8 -10.6 Example 18 100
73.9 .+-. 7.9 48.5 .+-. 4.9** -34.3 1000 91.7 .+-. 13.8 79.3 .+-.
17.0 -13.5 Example 23 300 93.5 .+-. 7.0 86.4 .+-. 10.0 -7.6 1000
74.6 .+-. 10.2 61.9 .+-. 8.5** -17.1 Example 42 300 83.1 .+-. 11.5
66.5 .+-. 9.9** -20.0 1000 77.4 .+-. 5.4 70.3 .+-. 7.8 -9.2 Example
51 300 80.0 .+-. 6.2 73.7 .+-. 6.1 -7.9 1000 78.3 .+-. 6.7 67.9
.+-. 5.6* -13.3 FLAVOXATE 300 89.2 .+-. 10.7 95.0 .+-. 10.9 +6.5
1000 90.4 .+-. 10.7 80.1 .+-. 11.1 -11.4 OXYBUTYNIN 100 95.2 .+-.
9.2 77.4 .+-. 10.3** -18.7 300 82.3 .+-. 8.7 50.5 .+-. 6.3** -38.6
*= P < 0.05, **= P < 0.01 versus basal values; Student's t
test for paired data
EXAMPLE 99
Binding to 5-HT.sub.1A and Other Different Neurotransmitter Binding
Sites
A. Methods
Recombinant Human 5HT 1A Receptors
[0260] Genomic clone G-21 coding for the human 5-HT1A serotonergic
receptor is stably transfected in a human cell line (HeLa). HeLa
cells were grown as monolayers in Dulbecco's modified Eagle's
medium (DMEM), supplemented with 10% fetal calf serum and
gentamicin (100 mg/ml), 5% CO.sub.2 at 37.degree. C. Cells were
detached from the growth flask at 95% confluence by a cell scraper
and were lysed in ice-cold 5 mM Tris and 5 mM EDTA buffer (pH 7.4).
Homogenates were centrifuged at 40000.times.g.times.20 min and
pellets were resuspended in a small volume of ice-cold 5 mM Tris
and 5 mM EDTA buffer (pH 7.4) and immediately frozen and stored at
-70.degree. C. until use. On the day of experiment, cell membranes
were resuspended in binding buffer: 50 mM Tris HCl (pH 7.4), 2.5 mM
MgCl2, 10 .mu.M pargiline (Fargin et al., Nature 335, 358-360,
1988). Membranes were incubated in a final volume of 1 ml for 30
min at 30.degree. C. with 0.2-1 nM [3H]8-OH-DPAT, in absence or
presence of competing drugs; non-specific binding was determined in
the presence of 10 .mu.M 5-HT. The incubation was stopped by
addition of ice-cold Tris-HCl buffer and rapid filtration through
0.2% polyethyleneimine pretreated Whatman GF/B or Schleicher &
Schuell GF52 filters.
Native 5-HT.sub.2A Serotoninergic Receptors and
.alpha..sub.2-adrenoceptor- s (from Animal Tissues)
[0261] Binding studies on native .alpha..sub.2 adrenergic receptors
(Diop L. et al, J. Neurochem. 41, 710-715, 1983), and 5-HT2A
serotonergic receptors (Craig A. and Kenneth J., Life Sci. 38,
117-127, 1986) were carried out in membranes of rat cerebral
cortex. Male Sprague Dawley rats (200-300 g, SD Harlan/Nossan,
Italy) were killed by cervical dislocation and cerebral cortexes
were excised and immediately frozen in liquid nitrogen and stored
at -70.degree. C. until use. Tissues were homogenized (2.times.20
sec) in 50 volumes of cold 50 mM Tris-HCl buffer pH 7.4, using a
Polytron homogenizer (speed 7). Homogenates were centrifuged at
49000.times.g for 10 min, resuspended in 50 volumes of the same
buffer, incubated at 37.degree. C. for 15 min and centrifuged and
resuspended twice more. The final pellets were suspended in 100
volumes of 50 mM Tris-HCl buffer pH 7.4, containing 10 .mu.M
pargiline and 0.1% ascorbic acid (a2 adrenergic receptors) or in
100 volumes of 50 mM Tris-HCl buffer pH 7.7 (5-HT2A serotonergic
receptors). Membranes were incubated in a final volume of 1 ml for
30 min at 25.degree. C. with 0.5-1.5 nM [3H]rauwolscine
(a2-adrenergic receptors) or for 20 min at 37.degree. C. with
0.7-1.3 nM [3H]ketanserin (5-HT2A receptors), in absence or
presence of competing drugs. Non-specific binding was determined in
the presence of 10 .mu.M phentolamine (a2-adrenergic receptors) or
2 .mu.M ketanserin (5-HT2A serotoninergic receptors). The
incubation was stopped by addition of ice-cold 50 mM Tris-HCl
buffer and rapid filtration through 0.2% polyethyleneimine
pretreated Whatman GF/B or Schleicher & Schuell GF52 filters.
The filters are then washed with ice-cold buffer and the
radioactivity retained on the filters was counted by liquid
scintillation spectrometry.
B. Results
[0262] The inhibition of specific binding of the radioligands by
the tested drugs was analyzed to estimate the IC50 value by using
the non-linear curve-fitting program Allfit (De Lean et al., Am. J.
Physiol. 235, E97-E102, 1978). The IC50 value was converted to an
affinity constant (Ki) by the equation of Cheng & Prusoff
(Cheng, Y. C.; Prusoff, W. H. Biochem. Pharmacol. 22, 3099-3108,
1973).
[0263] The results shown in Table 3A demonstrate that compound A
and the compound of Example 2 both have a very high affinity for
5-HT.sub.1A receptors, but their binding profile is different. The
compound of Example 2 was much more selective than compound A for
the 5-HT1A receptor versus the 5-HT.sub.2A and the
a2-adrenoceptors. All the other compounds of the invention tested
(Table 3B) had high affinity for the 5-HT1A receptor.
3TABLE 3B Binding affinity for the 5-HT.sub.1A receptor Data are
expressed as Ki (nM). Compound 5-HT.sub.1A Ex. 3 10.28 Ex. 4 0.64
Ex. 5 14.85 Ex. 6 0.45 Ex. 7 3.82 Ex. 8 0.36 Ex. 10 17.23 Ex. 11
2.92 Ex. 12 4.77 Ex. 13 0.50 Ex. 14 10.32 Ex. 15 6.20 Ex. 16 2.9
Ex. 17 20.15 Ex. 18 0.60 Ex. 20 24.62 Ex. 21 2.72 Ex. 22 18.18 Ex.
23 0.14 Ex. 25 8.91 Ex. 26 2.69 Ex. 27 0.57 Ex. 28 18.78 Ex. 30
7.96 Ex. 32 19.36 Ex. 34 16.27 Ex. 35 8.00 Ex. 38 1.02 Ex. 39 2.65
Ex. 45 2.21 Ex. 46 6.05 Ex. 47 2.33 Ex. 48 0.46 Ex. 49 0.17 Ex. 50
1.16 Ex. 51 0.24 Ex. 52 34 Ex. 53 11.75 Ex. 55 10.78 Ex. 56 2.13
Ex. 57 0.74 Ex. 58 2.44 Ex. 65 18.62 Ex. 66 0.76 Ex. 67 4.76 Ex. 68
9.32 Ex. 69 0.32 Ex. 70 28.09 Ex. 71 15.48 Ex. 72 1.22 Ex. 73 21.23
Ex. 74 2.02 Ex. 76 0.69 Ex. 77 0.58 Ex. 83 1.31 Ex. 84 23.23 Ex. 88
1.03 Ex. 93 1.60 Ex. 94 2.14 Ex. 96 1.20
Measurement of Pre- and Post-Synaptic 5-HT.sub.1A Receptor
Antagonist Activity
A. Methods
Antagonism of Hypothermia Induced by 8-OH-DPAT in Mice
(Pre-synaptic Antagonism).
[0264] The antagonistic effect of the .sup.5-HT.sub.1A receptor
antagonists of the invention on hypothermia induced by 8-OH-DPAT
was evaluated by the method of Moser (Moser, Eur. J. Pharmacol.,
193:165, 1991) with minor modifications as described below. Male
CD-1 mice (28-38 g) obtained from Charles River (Italy) were housed
in a climate-controlled room (temperature 22.+-.2 C.; humidity
55.+-.15%) and maintained on a 12 h light/dark cycle with free
access to food and water. On the day of experiment, mice were
placed singly in clear plastic boxes under the same ambient
conditions. Body temperature was measured by the insertion of a
temperature probe (Termist TM-S, LSI) into the rectum to a depth of
2 cm. Rectal temperature was measured immediately prior to
intravenous injection of the test compound. All animals then
received 8-OH-DPAT (0.5 mg/kg s.c.) and their temperature was
measured 30 min later. For each animal, temperature changes were
calculated with respect to pretreatment values and the mean values
were calculated for each treatment group. A linear regression
equation was used in order to evaluate ID.sub.50 values, defined as
the dose of antagonist needed to block 50% of the hypothermic
effect induced by 0.5 mg/kg 8-OH-DPAT administered
subcutaneously.
Inhibition Offorepaw Treading Induced by 8-OH-DPA Tin Rats
(Post-synaptic Antagonism)
[0265] The inhibitory effect of 5-HT.sub.1A receptor antagonists on
the forepaw treading induced in rats by subcutaneous injection of
8-OH-DPAT was evaluated by the method of Tricklebank (Tricklebank
et al., Eur. J. Pharmacol., 117:15, 1985) with minor modifications
as described below.
[0266] Male Sprague-Dawley rats (150-175 g) obtained from Charles
River (Italy), were housed in a climate-controlled room and
maintained on a 12 h light/dark cycle with free access to food and
water. On the day of experiment, rats were placed singly in clear
plastic boxes. Rats were treated with reserpine, 1 mg/kg s.c.,
18-24 h before the test to deplete intracellular stores of
noradrenaline. For evaluation of antagonistic activity, compounds
were i.v. administered 16 min before 8-OH-DPAT (1 mg/kg s.c.).
Observation sessions of 30 s duration began 3 min after treatment
with the agonist and were repeated every 3 min over a period of 15
min. The appearance of the forepaw treading symptom induced by
postsynaptic stimulation of the 5HT.sub.1A receptors was noted, and
its intensity was scored using a ranked intensity scale in which:
0=absent, 1=equivocal, 2=present and 3=intense. Behavioral scores
for each treated rat were accumulated over the time course (5
observation periods) and expressed as mean values of 8-10 rats. A
linear regression equation was used in order to evaluate ID.sub.50
values, defined as the dose of antagonist needed to block 50% of
the forepaw treading intensity induced by 1 mg/kg 8-OH-DPAT
administered subcutaneously.
B. Results
[0267] The results are shown in Table 4. These results demonstrate
that compound of Example 2 exhibits significant pre-synaptic and
post-synaptic 5-HT.sub.1A receptor antagonist activity. Compound A,
by contrast, proved at least 10 fold less active than compound of
Example 2 in both models.
4TABLE 4 Antagonistic activity for the pre- and post-synaptic
5-HT.sub.1A receptor. Data are expressed as ID.sub.50 in mg/kg.
Pre-synaptic 5-HT.sub.1A Post-synaptic 5-HT.sub.1A Compound
ID.sub.50 ID.sub.50 Compound A 221 350 Example 2 20 36 Example 13
-- 82 Example 18 n.a. 84 Example 23 -- 177
* * * * *